Identification of a two metastasis-related prognostic signature in the process of predicting the survival of laryngeal squamous cell carcinoma.

Metastasis is a major cause of treatment failure and poor outcomes in cancer patients. The data used in the current study was downloaded from TCGA and GEO databases. Differentially expressed metastasis-related genes were identified and the biological functions were implemented. Kaplan-Meier analysis univariate, and, multivariate Cox regression analyses were performed to identify robust prognostic biomarkers, followed by construction of the risk model and nomogram. Gene set enrichment analysis was performed to identify pathways enriched in low- and high-risk groups. POLR2J3 and MYH11 were treated as prognostic biomarkers in LSCC and the risk model was constructed. Receiver operating characteristic curves revealed the good performance of the risk model. A nomogram with high accuracy was constructed, as evidenced by calibration and decision curves. Moreover, we found that the expressions of POLR2J3 and MYH11 was significantly higher in metastasis tissues compared with those in non-metastasis tissues by RT-qPCR and IHC. Our study identified novel metastasis-related prognostic biomarkers in LSCC and constructed a unique nomogram for predicting the prognosis of LSCC patients. Moreover, we explored the related mechanisms of metastasis-related genes in regulating LSCC.

Clinical efficacy of styloid incision truncation via percutaneous punching in treating styloid process syndrome.

OBJECTIVE: To clarify the clinical efficacy of styloid incision truncation via percutaneous punching in treating styloid process (styloid) syndrome. METHODS: The clinical data of 40 styloid syndrome patients treated in our hospital from July 2018 to August 2021 were chosen and divided into an observation group and a control group in a random manner, with 20 cases in each. The control group received treatment with styloid truncation via an external cervical approach, and the observation group received treatment with styloid incision truncation via percutaneous punching. The operation time, intraoperative blood loss, length of truncated styloid, clinical efficacy, pain scores, postoperative complications and inflammatory cytokine levels were assessed in the both groups. RESULTS: The intraoperative blood loss, operation time, length of truncated styloid and hospital stay in the observation group were significantly lower than those in the control group (P < 0.05). VAS pain scores were higher in both groups after the operation compared to before the operation. However, the observation group showed a statistically significant reduction in comparison with the control group (P < 0.05). The treatment effectiveness and complication rates of the two groups exhibited significant differences (P < 0.05). After the operation, TNF-α, CRP, and IL-6 levels in both groups were elevated compared to those before the operation. The observation group, however, showed significant depletion compared to the control group (P < 0.05). CONCLUSION: Styloid incision truncation via percutaneous punching was not only effective in treating styloid syndrome, but also caused less trauma and fewer complications. It promotes patient recovery and requires a simple operation, making it worthy of promotion in hospitals.

Monitoring Therapeutic Responses to Silicified Cancer Cell Immunotherapy Using PET/MRI in a Mouse Model of Disseminated Ovarian Cancer.

Current imaging approaches used to monitor tumor progression can lack the ability to distinguish true progression from pseudoprogression. Simultaneous metabolic 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) positron emission tomography (PET) and magnetic resonance imaging (MRI) offers new opportunities to overcome this challenge by refining tumor identification and monitoring therapeutic responses to cancer immunotherapy. In the current work, spatial and quantitative analysis of tumor burden were performed using simultaneous [18F]FDG-PET/MRI to monitor therapeutic responses to a novel silicified cancer cell immunotherapy in a mouse model of disseminated serous epithelial ovarian cancer. Tumor progression was validated by bioluminescence imaging of luciferase expressing tumor cells, flow cytometric analysis of immune cells in the tumor microenvironment, and histopathology. While PET demonstrated the presence of metabolically active cancer cells through [18F]FDG uptake, MRI confirmed cancer-related accumulation of ascites and tissue anatomy. This approach provides complementary information on disease status without a confounding signal from treatment-induced inflammation. This work provides a possible roadmap to facilitate accurate monitoring of therapeutic responses to cancer immunotherapies.

Effects of Different Intervention Methods on Intestinal Cleanliness in Children Undergoing Colonoscopy.

Objective: To explore the effects of different intervention methods on intestinal cleanliness in children undergoing colonoscopy. Methods: 61 children who underwent colonoscopy in our hospital from May 2020 to May 2021 were randomly divided into group A (n = 21), group B (n = 30), and group C (n = 10). The children in the three groups were intervened in different ways before the colonoscopy. Group A received a long-handled Kaiselu +1 cathartic intervention, while group B received a long-handled Kaiselu +2 cathartic intervention, and group C received an enema plus one cathartic intervention. The patients in the three groups were given the same diet before the examination until the examination was completed. The time-related indexes, cleanliness, adverse reactions, tolerance, and adaptability of the three groups under different dietary interventions and cleaning methods were evaluated. Results: The first defecation time in group C was lower than that in group A and group B, the hospital stay was longer than that in group A and group B (p > 0.05), and the colonoscopy time in group C was shorter than that in group A and group B (p < 0.05). The BBPS score of group C was (2.10 ± 0.32), which was significantly higher than that of group A (1.16 ± 0.19) and group B (1.77 ± 0.18) (p < 0.05). The BBPS scores of children with liquid food in the three groups were significantly higher than those of common food, and the BBPS scores of liquid food and common food in group C were significantly higher than those in group A and group B (p < 0.05). The incidence of adverse reactions in group C was 20.00%, which was significantly lower than 33.33% in group A and 23.33% in group B (p < 0.05). The proportion of grade I in group C was 50.00%, which was significantly higher than 38.10% in group A and 43.33% in group B (p < 0.05). Conclusion: Children undergoing colonoscopy take preintestinal preparation under different diets and intervention methods. The cleanliness of liquid food and enema + one-time laxative one day before colonoscopy is the best, which can significantly reduce adverse reactions and increase the acceptability and adaptability of children. It is worthy of clinical application.

Circ_0023028 contributes to the progression of laryngeal squamous cell carcinoma by upregulating LASP1 through miR-486-3p.

Circular RNAs (circRNAs) are implicated in the tumorigenesis of human cancers. However, the effects of circRNAs on laryngeal squamous cell carcinoma (LSCC) are largely unknown. Here, we aimed to explore the roles of circ_0023028 in LSCC development. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure circ_0023028, miR-486-3p, and Lin-Isl-Mec (LIM) and SH3 domain protein 1 (LASP1) mRNA. The characteristics of circ_0023028 were determined by RNase R digestion assay and Actinomycin D assay. Cell Counting Kit-8 (CCK-8) assay and colony formation assay were utilized for cell proliferation. Transwell assay was adopted for cell invasion and migration. Flow cytometry analysis was carried out to analyze cell cycle and apoptosis. RNA pull-down assay and dual-luciferase reporter assay were used to explore the association between miR-486-3p and circ_0023028 or LASP1. Western blot assay was adopted to measure LASP1 protein level. Murine xenograft model was executed to investigate the function of circ_0023028 in vivo. High expression of circ_0023028 was observed in LSCC tissues and cells. Circ_0023028 was stable and possessed a loop structure. Circ_0023028 silencing suppressed LSCC cell proliferation, metastasis and cell cycle process and induced apoptosis in vitro and hampered tumor growth in vivo. Further mechanism analysis demonstrated that circ_0023028 could sponge miR-486-3p to regulate LASP1 expression in LSCC cells. The malignant behaviors of LSCC cells mediated by circ_0023028 knockdown were rescued by the inhibition of miR-486-3p. Moreover, miR-486-3p suppressed LSCC cell progression via binding to LASP1. Circ_0023028 knockdown might impede the progression of LSCC by regulating miR-486-3p/LASP1 axis, which could provide a novel insight on the mechanism of LSCC progression.

Kidney intercalated cells and the transcription factor FOXi1 drive cystogenesis in tuberous sclerosis complex.

Tuberous sclerosis complex (TSC) is caused by mutations in either TSC1 or TSC2 genes and affects multiple organs, including kidney, lung, and brain. In the kidney, TSC presents with the enlargement of benign tumors (angiomyolipomata) and cysts, which eventually leads to kidney failure. The factors promoting cyst formation and tumor growth in TSC are incompletely understood. Here, we report that mice with principal cell-specific inactivation of Tsc1 develop numerous cortical cysts, which are overwhelmingly composed of hyperproliferating A-intercalated (A-IC) cells. RNA sequencing and confirmatory expression studies demonstrated robust expression of Forkhead Transcription Factor 1 (Foxi1) and its downstream targets, apical H+-ATPase and cytoplasmic carbonic anhydrase 2 (CAII), in cyst epithelia in Tsc1 knockout (KO) mice but not in Pkd1 mutant mice. In addition, the electrogenic 2Cl-/H+ exchanger (CLC-5) is significantly up-regulated and shows remarkable colocalization with H+-ATPase on the apical membrane of cyst epithelia in Tsc1 KO mice. Deletion of Foxi1, which is vital to intercalated cells viability and H+-ATPase expression, completely abrogated the cyst burden in Tsc1 KO mice, as indicated by MRI images and histological analysis in kidneys of Foxi1/Tsc1 double-knockout (dKO) mice. Deletion of CAII, which is critical to H+-ATPase activation, caused significant reduction in cyst burden and increased life expectancy in CAII/Tsc1 dKO mice vs. Tsc1 KO mice. We propose that intercalated cells and their acid/base/electrolyte transport machinery (H+-ATPase/CAII/CLC-5) are critical to cystogenesis, and their inhibition or inactivation is associated with significant protection against cyst generation and/or enlargement in TSC.

Downregulation of miR­143 modulates KRAS expression in colorectal carcinoma cells.

MicroRNAs (miRs) are a class of non­coding small RNAs that have been demonstrated to be involved in the pathogenesis of human cancer. There is even evidence that microRNAs can act as oncogenes or tumor suppressors. Although microRNA expression profiles have been characterized in colorectal carcinoma, their precise physiological functions are largely unknown. It has become clear that the activated KRAS Proto­Oncogene, GTPase (KRAS) oncogene plays an important role in colorectal carcinogenesis. In the present study, it was found that the level of mature miR­143 was lower in colorectal carcinoma tissues compared with that observed in normal adjacent tissues. A lack of miR­143 was detected in human colorectal carcinoma cell lines, SW480, LoVo and HT­29, compared to the high expression observed in normal colon epithelial cell line NCM460. pcDNA3.1­pri­miR­143 and its mutant were successfully constructed and transfected into colorectal carcinoma cells. Increased accumulation of mature miR­143 was observed in the pcDNA3.1­pri­miR­143­transfected cells. In SW480 cells, transfection of pcDNA3.1­pri­miR­143 resulted in a 35 and 47% reduction in cell growth after incubation for 4 and 5 days, respectively, compared with transfection of the pcDNA3.1­pri­miR­143 mutant; while in LoVo cells, transfection of pcDNA3.1­pri­miR­143 resulted in a 33 and 46% reduction in cell growth respectively. In contrast, transfection of pcDNA3.1­pri­miR­143 had no significantly effects on HT­29 cell growth. We also found that transfection of pcDNA3.1­pri­miR­143 had no effect on levels of KRAS mRNA, but resulted in a 58% decrease in the KRAS protein level in the transfected SW480 cells, while an approximate 54 and 43% KRAS protein reduction in LoVo and HT­29 cells, respectively, compared with the pcDNA3.1­pri­miR­143 mutant. Two fragments containing the putative complementary site were cloned into the pGL3 vector, constructing the luciferase reporter pGL3­KRAS­CS1 and pGL3­KRAS­CS2. Cotransfection of pcDNA3.1­pri­miR­143 with pGL3­KRAS­CS1 and pGL3­KRAS­CS2 respectively resulted in 4.6­ and 3.3­fold inhibition of luciferase activity in the SW480 cells, while a 4.0­ and 3.2­fold inhibition of luciferase activity in the LoVo cells, 3.7­ and 3.1­fold inhibition in the HT­29 cells. Differences in pGL3­KRAS­CS1 and pGL3­KRAS­CS2 activity were not significant. Our results revealed that increased accumulation of miR­143 is likely to modulate levels of KRAS protein expression at the post­transcriptional level by interacting specifically with the complementary site, and consequently inhibiting proliferation of the transfected cells.

Microstructural and microglial changes after repetitive mild traumatic brain injury in mice.

Traumatic brain injury (TBI) is a major public health issue, with recently increased awareness of the potential long-term sequelae of repetitive injury. Although TBI is common, objective diagnostic tools with sound neurobiological predictors of outcome are lacking. Indeed, such tools could help to identify those at risk for more severe outcomes after repetitive injury and improve understanding of biological underpinnings to provide important mechanistic insights. We tested the hypothesis that acute and subacute pathological injury, including the microgliosis that results from repeated mild closed head injury (rmCHI), is reflected in susceptibility-weighted magnetic resonance imaging and diffusion-tensor imaging microstructural abnormalities. Using a combination of high-resolution magnetic resonance imaging, stereology, and quantitative PCR, we studied the pathophysiology of male mice that sustained seven consecutive mild traumatic brain injuries over 9 days in acute (24 hr) and subacute (1 week) time periods. rmCHI induced focal cortical microhemorrhages and impaired axial diffusivity at 1 week postinjury. These microstructural abnormalities were associated with a significant increase in microglia. Notably, microgliosis was accompanied by a change in inflammatory microenvironment defined by robust spatiotemporal alterations in tumor necrosis factor-α receptor mRNA. Together these data contribute novel insight into the fundamental biological processes associated with repeated mild brain injury concomitant with subacute imaging abnormalities in a clinically relevant animal model of repeated mild TBI. These findings suggest new diagnostic techniques that can be used as biomarkers to guide the use of future protective or reparative interventions. © 2016 Wiley Periodicals, Inc.

Imaging and serum biomarkers reflecting the functional efficacy of extended erythropoietin treatment in rats following infantile traumatic brain injury.

OBJECTIVE Traumatic brain injury (TBI) is a leading cause of death and severe morbidity for otherwise healthy full-term infants around the world. Currently, the primary treatment for infant TBI is supportive, as no targeted therapies exist to actively promote recovery. The developing infant brain, in particular, has a unique response to injury and the potential for repair, both of which vary with maturation. Targeted interventions and objective measures of therapeutic efficacy are needed in this special population. The authors hypothesized that MRI and serum biomarkers can be used to quantify outcomes following infantile TBI in a preclinical rat model and that the potential efficacy of the neuro-reparative agent erythropoietin (EPO) in promoting recovery can be tested using these biomarkers as surrogates for functional outcomes. METHODS With institutional approval, a controlled cortical impact (CCI) was delivered to postnatal Day (P)12 rats of both sexes (76 rats). On postinjury Day (PID)1, the 49 CCI rats designated for chronic studies were randomized to EPO (3000 U/kg/dose, CCI-EPO, 24 rats) or vehicle (CCI-veh, 25 rats) administered intraperitoneally on PID1-4, 6, and 8. Acute injury (PID3) was evaluated with an immunoassay of injured cortex and serum, and chronic injury (PID13-28) was evaluated with digitized gait analyses, MRI, and serum immunoassay. The CCI-veh and CCI-EPO rats were compared with shams (49 rats) primarily using 2-way ANOVA with Bonferroni post hoc correction. RESULTS Following CCI, there was 4.8% mortality and 55% of injured rats exhibited convulsions. Of the injured rats designated for chronic analyses, 8.1% developed leptomeningeal cyst-like lesions verified with MRI and were excluded from further study. On PID3, Western blot showed that EPO receptor expression was increased in the injured cortex (p = 0.008). These Western blots also showed elevated ipsilateral cortex calpain degradation products for αII-spectrin (αII-SDPs; p < 0.001), potassium chloride cotransporter 2 (KCC2-DPs; p = 0.037), and glial fibrillary acidic protein (GFAP-DPs; p = 0.002), as well as serum GFAP (serum GFAP-DPs; p = 0.001). In injured rats multiplex electrochemiluminescence analyses on PID3 revealed elevated serum tumor necrosis factor alpha (TNFα p = 0.01) and chemokine (CXC) ligand 1 (CXCL1). Chronically, that is, in PID13-16 CCI-veh rats, as compared with sham rats, gait deficits were demonstrated (p = 0.033) but then were reversed (p = 0.022) with EPO treatment. Diffusion tensor MRI of the ipsilateral and contralateral cortex and white matter in PID16-23 CCI-veh rats showed widespread injury and significant abnormalities of functional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD); MD, AD, and RD improved after EPO treatment. Chronically, P13-P28 CCI-veh rats also had elevated serum CXCL1 levels, which normalized in CCI-EPO rats. CONCLUSIONS Efficient translation of emerging neuro-reparative interventions dictates the use of age-appropriate preclinical models with human clinical trial-compatible biomarkers. In the present study, the authors showed that CCI produced chronic gait deficits in P12 rats that resolved with EPO treatment and that chronic imaging and serum biomarkers correlated with this improvement.

Attenuation of acute stroke injury in rat brain by minocycline promotes blood-brain barrier remodeling and alternative microglia/macrophage activation during recovery.

BACKGROUND: Minocycline reduces reperfusion injury by inhibiting matrix metalloproteinases (MMPs) and microglia activity after cerebral ischemia. Prior studies of minocycline investigated short-term neuroprotective effects during subacute stage of stroke; however, the late effects of minocycline against early reperfusion injury on neurovascular remodeling are less well studied. We have shown that spontaneous angiogenesis vessels in ischemic brain regions have high blood-brain barrier (BBB) permeability due to lack of major tight junction proteins (TJPs) in endothelial cells at three weeks. In the present study, we longitudinally investigated neurological outcome, neurovascular remodeling and microglia/macrophage alternative activation after spontaneous and minocycline-induced stroke recovery. METHODS: Adult spontaneously hypertensive rats had a 90 minute transient middle cerebral artery occlusion. At the onset of reperfusion they received a single dose of minocycline (3 mg/kg intravenously) or a vehicle. They were studied at multiple time points up to four weeks with magnetic resonance imaging (MRI), immunohistochemistry and biochemistry. RESULTS: Minocycline significantly reduced the infarct size and prevented tissue loss in the ischemic hemispheres compared to vehicle-treated rats from two to four weeks as measured with MRI. Cerebral blood flow measured with arterial spin labeling (ASL) showed that minocycline improved perfusion. Dynamic contrast-enhanced MRI indicated that minocycline reduced BBB permeability accompanied with higher levels of TJPs measured with Western blot. Increased MMP-2 and -3 were detected at four weeks. Active microglia/macrophage, surrounding and within the peri-infarct areas, expressed YM1, a marker of M2 microglia/macrophage activation, at four weeks. These microglia/macrophage expressed both pro-inflammatory factors tumor necrosis factors-α (TNF-α) and interleukin-1ß (IL-1ß) and anti-inflammatory factors transforming growth factor-ß (TGF-ß) and interleukin-10 (IL-10). Treatment with minocycline significantly reduced levels of TNF-α and IL-1ß, and increased levels of TGF-ß, IL-10 and YM1. CONCLUSIONS: Early minocycline treatment against reperfusion injury significantly promotes neurovascular remodeling during stroke recovery by reducing brain tissue loss, enhancing TJP expression in ischemic brains and facilitating neuroprotective phenotype alternative activation of microglia/macrophages.

Simultaneous monitoring of CMV and BKV by quantitative PCR in renal transplant recipients.

Polyomavirus (BKV) and cytomegalovirus (CMV) are associated with renal graft failure. The aim was to establish a quantitative PCR method (Q-PCR) to detect BKV and CMV simultaneously. The conserved sequences of BKV and CMV were amplified and cloned into the plasmids as standards. The sensitivity, specificity and the precision of the assay were evaluated. Q-PCR was used to detect BKV and CMV DNA simultaneously in 480 blood samples of renal transplantation recipients. The sensitivity of the Q-PCR assay to detect BKV or CMV DNA reached 5×10(3)copies/mL. The use of control DNA verified that the assay could specifically detect the target DNA. The precision of the assay to quantify target DNA copies was acceptable (ICV 3.44% for BKV and 2.23% for CMV; differences between batches ICV 4.98% for BKV and 3.76% for CMV). In 480 samples, 130 samples (27.08%) were CMV DNA positive, which was significantly higher than the 64 BKV DNA positive samples (13.33%, p<0.05). BKV or CMV DNA positivity was significantly associated with high concentrations of Tacrolimus (TAC) (p value<0.05). The Q-PCR assay to detect both CMV and BKV DNA simultaneously was developed successfully with high sensitivity, precision, and time-effectiveness for clinical measurement.

The progression of the tubulointerstitial fibrosis driven by stress-induced "proliferation-death" vicious circle.

Several hypotheses have been developed to interpret the progression of tubulointerstitial fibrosis (TF), including senescence, epithelial-mesenchymal transition, inflammation, chronic hypoxia, and reactive oxygen species. All of these hypotheses are based on persistent cell injury and localized cell death. Proliferation of neighboring renal tubular epithelial cells (RTECs) is beneficial for organ function recovery from acute injury. However, compensatory proliferation is not always advantageous, as the proliferating cells are vulnerable to ongoing detrimental stimuli, such as inflammation, endocrine stress, high blood pressure, hypoxia/ischemia, and the like. Cell injury and death promotes secretion of growth factors, which evokes proliferation of RTECs; entering the cell cycle makes the RTECs more vulnerable to injury and death. Under persistent stress, death and proliferation are mutually promoted and form the vicious circle that triggers, maintains, and augments the inflammation and progression of TF. We hypothesize that the "proliferation-death" circle is another important pathophysiologic mechanism of TF onset. Through this hypothesis, this paper interprets the development and progression of TF. Moreover, the vicious circle may be universal, underlying the development of inflammation and fibrosis in various organs and tissues. The hypothesis also suggests a potential therapy strategy for the inhibition of fibrosis.

Effect of pulsed electromagnetic field (PEMF) on infarct size and inflammation after cerebral ischemia in mice.

Pulsed electromagnetic fields (PEMF) have been demonstrated to have anti-inflammatory and pro-regenerative effects in animals and humans. We used the FDA-approved Sofpulse (Ivivi Health Sciences, LLC) to study effect of PEMF on infarct size and poststroke inflammation following distal middle cerebral artery occlusion (dMCAO) in mice. Electromagnetic field was applied within 30-45 min after ischemic brain damage and utilized twice a day for 21 consecutive days. Ischemic infarct size was assessed using MRI and histological analysis. At 21 days after dMCAO, the infarct size was significantly (by 26%) smaller in PEMF-treated animals as compared to controls. Neuroinflammation in these animals was evaluated using specialized cytokine/chemokine PCR array. We demonstrate that PEMF significantly influenced expression profile of pro- and anti-inflammatory factors in the hemisphere ipsilateral to ischemic damage. Importantly, expression of gene encoding major pro-inflammatory cytokine IL-1α was significantly reduced, while expression of major anti-inflammatory IL-10 was significantly increased. PEMF application significantly downregulated genes encoding members of the major pro-apoptotic tumor necrosis factor (TNF) superfamily indicating that the treatment could have both anti-inflammatory and anti-apoptotic effects. Both reduction of infarct size and influence on neuroinflammation could have a potentially important positive impact on the poststroke recovery process, implicating PEMF as a possible adjunctive therapy for stroke patients.

Development of a nuclear morphometric signature for prostate cancer risk in negative biopsies.

BACKGROUND: Our objective was to develop and validate a multi-feature nuclear score based on image analysis of direct DNA staining, and to test its association with field effects and subsequent detection of prostate cancer (PCa) in benign biopsies. METHODS: Tissue sections from 39 prostatectomies were Feulgen-stained and digitally scanned (400×), providing maps of DNA content per pixel. PCa and benign epithelial nuclei were randomly selected for measurement of 52 basic morphometric features. Logistic regression models discriminating benign from PCa nuclei, and benign from malignant nuclear populations, were built and cross-validated by AUC analysis. Nuclear populations were randomly collected <1 mm or >5 mm from cancer foci, and from cancer-free prostates, HGPIN, and PCa Gleason grade 3-5. Nuclei also were collected from negative biopsy subjects who had a subsequent diagnosis of PCa and age-matched cancer-free controls (20 pairs). RESULTS: A multi-feature nuclear score discriminated cancer from benign cell populations with AUCs of 0.91 and 0.79, respectively, in training and validation sets of patients. In prostatectomy samples, both nuclear- and population-level models revealed cancer-like features in benign nuclei adjacent to PCa, compared to nuclei that were more distant or from PCa-free glands. In negative biopsies, a validated model with 5 variance features yielded significantly higher scores in cases than controls (P = 0.026). CONCLUSIONS: A multifeature nuclear morphometric score, obtained by automated digital analysis, was validated for discrimination of benign from cancer nuclei. This score demonstrated field effects in benign epithelial nuclei at varying distance from PCa lesions, and was associated with subsequent PCa detection in negative biopsies. IMPACT: This nuclear score shows promise as a risk predictor among men with negative biopsies and as an intermediate biomarker in Phase II chemoprevention trials. The results also suggest that subvisual disturbances in nuclear structure precede the development of pre-neoplastic lesions.

Cordyceps sinensis extracts attenuate aortic transplant arteriosclerosis in rats.

BACKGROUND: Transplant arteriosclerosis is a hallmark of chronic rejection and is still the major limiting factor affecting the success of long-term organ transplants. Development of transplant arteriosclerosis is refractory to conventional immunosuppressive drugs, and adequate therapy is not yet available. The aim of this study was to determine the role of Cordyceps sinensis extracts in reducing the formation of transplant arteriosclerosis in a rat aortic transplant model. METHODS: Lewis rat aortic allografts were transplanted into Brown-Norway recipient rats. Recipients received 0.5, 1, 2, and 5 mg/kg of Cordyceps sinensis extracts (or control saline) daily via intragastric injection for 60 d. Grafts were harvested 60 d post-transplantation and intimal thickness determined microscopically following hematoxylin and eosin (H and E) staining and abdominal aorta protein profiles determined by Western blot analysis. Cellular localization was assessed by histology and immunohistochemistry and the serum analyzed for tumor necrosis factor alpha (TNF-α) and intercellular adhesion molecule-1 (ICAM-1) by enzyme-linked immunosorbent assay (ELISA). RESULTS: C. sinensis administration resulted in a significant reduction in neointimal formation (neointimal thickness 8.27 ± 1.95 µm [0.5 mg/kg], 3.69 ± 1.43 µm [1 mg/kg], 3.69 ± 1.43 µm [1 mg/kg], 3.69 ± 1.43 µm [1 mg/kg] versus 11.42 ± 2.67 µm [control]) and in the proliferative activity of vascular smooth muscle cells. In addition, localized expression of TNF-α and ICAM-1 in transplant aortas was characterized by immunohistochemistry and immunoblot analyses demonstrating that C. sinensis treatment significantly reduced TNF-α and ICAM-1 levels compared with levels observed in controls (P < 0.05). Serum TNF-α and ICAM-1 levels were significantly reduced in C. sinensis-treated animals compared with controls (P < 0.05). CONCLUSION: C. sinensis treatment effectively reduced the formation of transplant arteriosclerosis in a rat aortic transplant model.

[Bacterial 16S rRNA genes and expression of IL-1ß, TNF-α and IgA in prostate tissues].

OBJECTIVE: To investigate the role of bacteria in the etiology of chronic prostatitis. METHODS: Complete prostate specimens were obtained at autopsy from 192 organ donors (aged 20 - 38 years old) during 2002 to 2008 who died of non-prostatic diseases. One tissue taken from the peripheral prostatic zone according to McNeal was divided into two pieces. One piece of tissue was taken for routine pathological examinations and immunohistochemical studies of interleukin (IL)-1ß, tumor necrosis factor-α (TNF-α) and IgA. Another one was taken for PCR assay to detect the bacterial 16S rRNA genes (16S rDNA). RESULTS: Of 192 prostate specimens, 64 (33.3%) had pathological changes of chronic prostatitis and 38 (19.8%) specimens was positive for bacterial 16S rDNA. Positive rates of 16S rDNA in chronic prostatitis and non-prostatitis specimens were 50.0% (32/64) and 4.6% (6/128) respectively (χ(2) = 55.185, P < 0.001). Expressions of IL-1ß, TNF-α and IgA in specimens of chronic prostatitis were significantly higher than those in non-prostatitis specimens (P < 0.001). A positive correlation could be found among three immunohistochemical indicators (P < 0.01). In 64 specimens with chronic prostatitis, a significant expression of IL-1ß, TNF-α and IgA was more often demonstrated in 16S rDNA positive group than in 16S rDNA negative group (P < 0.001). CONCLUSIONS: The up-regulations of bacterial 16S rDNA, cytokines and immunoglobulin A are involved in inflammatory response of chronic prostatitis. Bacterial infection may be an important cause of chronic prostatitis.

[Correlation of HSP90 mRNA expression with migration ability of human multiple myeloma cells].

This study was aimed to investigate the correlation of heat shock protein 90 (HSP90) expression with migration ability of human multiple myeloma cells. The HSP90 mRNA expression and migration change of human multiple myeloma cell line (U266) were detected by RT-PCR and Transwell chamber respectively after treatment of U266 cells with final concentration 50, 100, 150, 200 nmol/L of bortezomib (proteosome inhibitor) for 4 hours. The results indicated that along with the increasing of bortezomib concentration, the expression level of HSP90alpha mRNA in U266 cells was enhanced, while no obvious increase of HSP90beta mRNA expression was observed in spite of statistical difference as a whole (p<0.05), but with the increasing of drug concentration in cells, their migration ability gradually decreased (p<0.05). It is concluded that the correlation of HSP90 expression with migration ability of human multiple myeloma cells exists.

[Expressions of bacterial 16S rRNA, IL-1beta, TNF-alpha and NGF in prostate tissues].

OBJECTIVE: To investigate the role of bacteria in the etiology of chronic prostatitis. METHODS: A total of 162 complete prostate specimens were obtained at autopsy from organ donors (aged 20 -38 yr) who died of non-prostatic diseases. Each of the samples from the peripheral zone of the prostate was divided into two parts, one for routine pathological examination and immunohistochemical studies of interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF-alpha) and the nerve growth factor (NGF), and the other for PCR assay to detect the bacterial 16S rRNA gene (16S rDNA). RESULTS: Fifty-one (31.5%) of the total specimens presented pathological changes of chronic prostatitis, of which 44 had mild focal stromal, 5 mild focal stromal and periglandular and 2 mild focal periglandular inflammation. The positive rate of 16S rDNA was 19.1% (31/162), 51.0% (26/51) in the chronic prostatitis and 4.5% (5/111) in the non-prostatitis specimens (chi2 = 29.783, P < 0.01). In the specimens with chronic prostatitis, the expressions of IL-1beta, TNF-alpha and NGF were significantly higher in the 16S rDNA positive than in the 16S rDNA negative group (P < 0.01). CONCLUSION: Bacterial inflammation may play an important role in the etiology of chronic prostatitis.

[Mechanism of C-reactive protein on proliferation of multiple myeloma U266 cells].

This study was purposed to investigate the mechanism of C-reactive protein (CRP) on proliferation of U266 cells. The human multiple myeloma cell line U266 was incubated with human CRP (0, 5, 10, 20 mg/L) for 24 hours, then the proliferation level of U266 cells was detected by using blood analyser. The mRNA expressions of survivin and HSP90alpha were examined by RT-PCR. The results showed that the proliferation ratio was increased, as compared with the control group (p<0.05); furthermore, the mRNA levels of survivin and HSP90alpha were up-regulated in proportion to the increased CRP concentrations. There was significant correlation between expression of survivin and HSP90alpha (r=0.737, p<0.0001) in incubated cells. It is concluded that CRP can stimulate the proliferation of MM cells directly by up-regulating the expression of survivin and HSP90alpha in MM cells. CRP can be regarded as a potential target for MM treatment.

A fast SSP-PCR method for genotyping the ATP-binding cassette subfamily B member 1 gene C3435T and G2677T polymorphisms in Chinese transplant recipients.

AIM: P-glycoprotein, the product of the ATP-binding cassette subfamily B member 1 (ABCB1) gene (or the so-called multidrug resistance 1 gene), is an ATP-driven efflux pump contributing to the pharmacokinetics as well as the pharmacokinetics of drugs that are P-glycoprotein substrates, such as tacrolimus. This paper describes the development of a new method for detection of the 3435C/T and 2677G/T/A single nucleotide polymorphisms of the ABCB1 gene. The method is a simple sequence-specific primer polymerase chain reaction (SSP-PCR). METHODS: 158 Chinese health checkup examinees and 214 transplant recipients were included in the study. Genomic DNA was extracted from peripheral blood and amplified with SSP-PCR to detect the 3435C/T and 2677G/T/A mutations in ABCB1. The SSP-PCR condition was optimized, and the PCR results were compared with those of DNA sequencing. RESULTS: In the optimized condition, the two polymorphisms could be clearly distinguished after one-step PCR and electrophoresis. The ABCB1 3435C/T and 2677G/T/A genotypes of the subjects were scanned, and allele-specific bands were successfully amplified by SSP-PCR, which were in full accordance with the results of sequencing. CONCLUSION: As a fast, simple and inexpensive genotyping tool, the method would be practicable in large clinical studies on interindividual pharmacokinetics.