[The influence of knocking down the expression of low-density lipoprotein receptor associated proteins on the vascular abnormalities in hepatocellular carcinoma and its mechanisms].

Objectives: To investigate the effect of the expression of low-density lipoprotein receptor associated protein (LDLR) on the vascular abnormalities in hepatocellular carcinoma (HCC) and its mechanisms. Methods: Based on the information of Oncomine Cancer GeneChip database, we analyzed the correlation between the expression level of LDLR and the expression level of carcinoembryonic antigen (CEA) and CD31 in hepatocellular carcinoma tissues. Lentiviral transfection of short hairpin RNA target genes was used to construct LDLR-knockdown MHCC-97H and HLE hepatocellular carcinoma cells. The differential genes and their expression level changes in LDLR-knockdown hepatocellular carcinoma cells were detected by transcriptome sequencing, real-time fluorescence quantitative polymerase chain reaction, and protein immunoblotting. The gene-related signaling pathways that involve LDLR were clarified by enrichment analysis. The effect of LDLR on CEA was assessed by the detection of CEA content in conditioned medium of hepatocellular carcinoma cells. Angiogenesis assay was used to detect the effect of LDLR on the angiogenic capacity of human umbilical vein endothelial cells, as well as the role of CEA in the regulation of angiogenesis by LDLR. Immunohistochemical staining was used to detect the expression levels of LDLR in 176 hepatocellular carcinoma tissues, and CEA and CD31 in 146 hepatocellular carcinoma tissues, and analyze the correlations between the expression levels of LDLR, CEA, and CD31 in the tissues, serum CEA, and alanine transaminase (ALT). Results: Oncomine database analysis showed that the expressions of LDLR and CEA in the tissues of hepatocellular carcinoma patients with portal vein metastasis were negatively correlated (r=-0.64, P=0.001), whereas the expressions of CEA and CD31 in these tissues were positively correlated ( r=0.46, P=0.010). The transcriptome sequencing results showed that there were a total of 1 032 differentially expressed genes in the LDLR-knockdown group and the control group of MHCC-97H cells, of which 517 genes were up-regulated and 515 genes were down-regulated. The transcript expression level of CEACAM5 was significantly up-regulated in the cells of the LDLR-knockdown group. The Gene Ontology (GO) function enrichment analysis showed that the differential genes were most obviously enriched in the angiogenesis function. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis showed that the relevant pathways involved mainly included the cellular adhesion patch, the extracellular matrix receptor interactions, and the interactions with the extracellular matrix receptors. The CEA content in the conditioned medium of the LDLR-knockdown group was 43.75±8.43, which was higher than that of the control group (1.15±0.14, P＜0.001). The results of angiogenesis experiments showed that at 5 h, the number of main junctions, the number of main segments, and the total area of the lattice formed by HUVEC cells cultured with the conditioned medium of MHCC-97H cells in the LDLR-knockdown group were 295.3±26.4, 552.5±63.8, and 2 239 781.0±13 8211.9 square pixels, which were higher than those of the control group (113.3±23.5, 194.8±36.5, and 660 621.0±280 328.3 square pixels, respectively, all P＜0.01).The number of vascular major junctions, the number of major segments, and the total area of the lattice formed by HUVEC cells cultured in conditioned medium with HLE cells in the LDLR-knockdown group were 245.3±42.4, 257.5±20.4, and 2 535 754.5±249 094.2 square pixels, respectively, which were all higher than those of the control group (113.3±23.5, 114.3±12.2, and 1 565 456.5±219 259.7 square pixels, respectively, all P＜0.01). In the conditioned medium for the control group of MHCC-97H cells,the number of main junctions, the number of main segments, and the total area of the lattice formed by the addition of CEA to cultured HUVEC cells were 178.9±12.0, 286.9±12.3, and 1 966 990.0±126 249.5 spixels, which were higher than those in the control group (119.7±22.1, 202.7±33.7, and 1 421 191.0±189 837.8 square pixels, respectively). The expression of LDLR in hepatocellular carcinoma tissues was not correlated with the expression of CEA, but was negatively correlated with the expression of CD31 (r=-0.167, P=0.044), the level of serum CEA (r=-0.061, P=0.032), and the level of serum ALT(r=-0.147,P=0.05). The expression of CEA in hepatocellular carcinoma tissues was positively correlated with the expression of CD31 (r=0.192, P=0.020). The level of serum CEA was positively correlated with the level of serum ALT (r=0.164, P=0.029). Conclusion: Knocking down LDLR can promote vascular abnormalities in HCC by releasing CEA.

[Updates and advances in the concept of gallbladder cancer treatment].

The gallbladder is the most common site of tumor occurrence among biliary tract cancer. Gallbladder cancer accounts for approximately 0.6% of new cancers and 0.9% of cancer-related deaths. The risk factors identified for the development of gallbladder cancer include being female,>65 years of age, asymptomatic gallstone disease,and obesity. Surgical resection is the only curative treatment for early-stage gallbladder cancer, and some intermediate or advanced gallbladder cancers can be radically cured by extended resection. However, the extent of liver resection or lymph node dissection and whether to combine it with bile duct removal, revascularisation,and multiple organ resection remain somewhat controversial. After neoadjuvant treatment, up to a third of patients with locally advanced gallbladder cancer benefit from secondary surgical treatment. Only a small proportion of patients with gallbladder cancer at high risk for recurrence will benefit from postoperative adjuvant therapy. With the advent of different target-targeted drugs and the use of genetic tests in biliary tract cancer, targeted therapy and PD-1/PD-L1 inhibitors may become the new standard of care for gallbladder cancer and need to be further explored.

[Implantation of a monofocal intraocular lens after femtosecond laser intrastromal keratotomy and contralateral implantation of a trifocal IOL: a case report].

An age-related cataract patient who underwent femtosecond laser intrastromal keratotomy in the right eye for presbyopia correction 8 years ago was subjected to femtosecond laser-assisted phacoemulsification, with implantation of a monofocal intraocular lens (IOL) and a trifocal IOL in the right and left eyes, respectively. The corneal stromal ring was complementary to the monofocal IOL, which recovered the distance and near visual acuity, in the right eye postoperatively. The trifocal IOL provided good intermediate visual acuity for the left eye. The vision of the patient reached an ideal level for all visual distances. The binocular fusion was within the normal range, and the stereoscopic vision was restored. We hope that this case report can act as a reference for the treatment of cataract after presbyopia corrective surgery.

Current locoregional therapies and treatment strategies in hepatocellular carcinoma.

Locoregional therapies (lrts) play an important role in the treatment of hepatocellular carcinoma (hcc), with the aim of increasing overall survival while preserving liver function. Various forms of lrt are available, and choosing the best one depends on technical aspects, liver morphology, tumour biology, and the patient's symptoms. The purpose of the present review article is to provide an overview of the current evidence relating to the use of percutaneous ablation, transarterial chemoembolization, and transarterial radioembolization for the curative or palliative treatment of hcc. Special situations are also reviewed, including the combined use of systemic therapy and lrt, indications and techniques for bridging to transplant and downstaging, and the use of lrt to treat patients with hcc and macrovascular invasion.

Optimized nerve block techniques while performing percutaneous hepatic ablation: Literature review and practical use.

Percutaneous image guided thermal ablation has become a cornerstone of therapy for patients with oligometastatic disease and primary liver malignancies. Evolving from percutaneous ethanol injection (PEI), thermal ablation utilizing radiofrequency ablation (RFA) and microwave ablation (MWA) have become the standard approach in the treatment of isolated lesions that fit within the size criteria for curative intent therapy (typically 3-4cm). With the evolution of more intense thermal ablation, such as MWA, the dramatic increase in both the size of ablation zone and intensity of heat generation have extended the limits of this technique. As a result of these innovations, intra-procedural and post-procedural pain have also significantly increased, requiring either higher levels of intravenous sedation or, in some institutions, general anesthesia. In addition to the increase in therapeutic intensity, the use of intravenous sedation during aggressive ablation procedures carries the risk of over-sedation when the noxious insult (i.e. the ablation) is removed, adding further difficulty to post-procedural recovery and management. Furthermore, high subdiaphragmatic lesions become challenging in this setting due to issues relating to sedation and compliance with breath hold/breathing instructions. Although general anesthesia may mitigate these complications, the added resources associated with providing general anesthesia during ablation is not cost effective and may result in substantial delays in treatment. The reduction of Aerosol Generating Medical Procedures (AGMP), such as intubation due to the COVID-19 Pandemic, must also be taken into consideration. Due to the potential increased risk of infection transmission, alternatives to general anesthesia should be considered when safe and possible. Upper abdominal regional nerve block techniques have been used to manage pain related to trauma, surgery, and cancer; however, blocks of this nature are not well described in the interventional radiology literature. The McGill University group has developed experience in using such blocks as splanchnic, celiac and hepatic hilar nerve blocks to provide peri-procedural pain control [1]. Since incorporating these techniques (along with hydrodissection with tumescent anesthesia), we have also observed in our high volume ablation center a dramatic decrease in the amount of sedatives administered during the procedure, a decrease in patient discomfort during localization and ablation, as well as decreased pain post-procedure. Faster time to discharge and overall reduction in room procedural time serve as added benefits. The purpose of this publication is to outline and illustrate the practical application and use of nerve block/regional anesthesia techniques with respect to percutaneous hepatic thermal ablation.

Hsa-miR-375 promotes the progression of inflammatory bowel disease by upregulating TLR4.

OBJECTIVE: To elucidate the biological function of hsa-miR-375 in the progression of inflammatory bowel disease (IBD) and the potential mechanism. PATIENTS AND METHODS: Intestinal mucosa tissues of 26 IBD patients and 30 healthy volunteers who underwent colonoscopy were harvested for determining hsa-miR-375 level by quantitative Real-time polymerase chain reaction (qRT-PCR). Binding of hsa-miR-375 to toll-like receptor 4 (TLR4) was verified by the dual-luciferase reporter gene assay. Changes in the viability and apoptosis in Caco-2 cells influenced by hsa-miR-375 were examined by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. The regulatory effect of hsa-miR-375 on the intestinal epithelial barrier was examined by detecting transepithelial electrical resistance (TEER) and lucifer yellow flux. Relative levels of TLR4, nuclear factor-kappa B (NF-κB), zonula occludens-1 (ZO-1), occludin and inflammatory factors in Caco-2 cells were detected by qRT-PCR, Western blot and enzyme-linked immunosorbent assay (ELISA). RESULTS: Hsa-miR-375 was downregulated in intestinal mucosa tissues of patients with Crohn's disease (CD) and ulcerative colitis (UC). Knockdown of hsa-miR-375 decreased viability and TEER, but elevated apoptotic rate and lucifer yellow flux. Overexpression of hsa-miR-375 achieved the opposite trends. TLR4 was the direct downstream of hsa-miR-375, and its level was negatively mediated by hsa-miR-375. In addition, TLR4 level in Caco-2 cells was upregulated after LPS induction, while hsa-miR-375 level was unchangeable. Knockdown of hsa-miR-375 upregulated NF-κB and pro-inflammatory factors TNF-α, IL-1ß, IL-6 and IL-8, and downregulated ZO-1, occludin and anti-inflammatory factor IL-10. CONCLUSIONS: Hsa-miR-375 is involved in the pathogenesis of IBD by upregulating TLR4 and inducing NF-κB activation.

Decreased expression of miR-551b predicts poor prognosis and promotes tumorigenesis by targeting PTP4A3 in human colorectal cancer.

OBJECTIVE: MiR-551b has been reported to display tumor-suppressive and oncogenic potential in several cancers, but there has been no study on the roles of miR-551b in colorectal cancer (CRC). In this work, we aimed to explore the potential functions and mechanisms of miR-551b in the modulation of the CRC progression. PATIENTS AND METHODS: The expressions of miR-551b were examined in 122 pairs of CRC cancer tissues and adjacent non-tumor samples by Real Time-Polymerase Chain Reaction (RT-PCR). The clinical significance of miR-551b in CRC patients was explored using a Chi-square test, Kaplan-Meier assays, and multivariate analysis. MiR-551b mimics and inhibitors were used to establish miR-551b upregulation and downregulation models in CRC cells to examine the functions of miR-551b on cells proliferation, migration, invasion, and apoptosis. Dual-luciferase reporter assays were conducted for the validation of the possible modulation of a putative target of miR-551b. RESULTS: We showed that miR-551b was significantly down-regulated in CRC tissues and cell lines. It was observed that miR-551b expressions were correlated with lymph nodes metastasis, TNM stage, and poor prognosis. Multivariate analysis identifies low level of miR-551b expressions as an independent predictor for a shorter overall survival. The functional assessment suggested that forced miR-551b expression distinctly suppressed CRC cells growth, invasion, and migration, while the suppression of miR-551b displayed the opposite trend. Mechanistic studies confirmed that PTP4A3 was identified as a direct target of miR-551b in CRC. Interesting observations revealed that the cells capacities were higher in miR-551b +PTP4A3 group, when compared with those in miR-551b group, indicating that up-regulation of PTP4A3 rescued the repressive functions of miR-551b overexpression on CRC cells growth and migration. CONCLUSIONS: The findings of our study first showed that miR-551b, a potential tumor suppressor, may be used as a promising prognostic predictor and therapeutic target for CRC.

Has_circ_0008274 promotes cell proliferation and invasion involving AMPK/mTOR signaling pathway in papillary thyroid carcinoma.

OBJECTIVE: Circular RNAs (circRNAs) have been known as important regulators in tumorigenesis. Whether circRNAs are involved in papillary thyroid carcinoma (PTC) requires to be determined. In the present study, we aimed to investigate the expression and function of has_circ_0008274 in PTC. PATIENTS AND METHODS: Tissue expression of has_circ_0008274 was evaluated in Gene Expression Omnibus datasets (GSE93522). Real-time PCR assays were used to detect the expression of has_circ_0008274 in human PTC tissues and cell lines. The correlation of has_circ_0008274 expression with clinicopathological factors was statistically analyzed. The MTT assay, colony formation assay, transwell assays were performed to analyze and compare cell viability and invasion. Western blot analysis was used to quantify the expression of AMPK/mTOR signaling pathway proteins. RESULTS: We found that has_circ_0008274 was significantly upregulated in PTC tissues, and the level of has_circ_0008274 was negatively associated with TNM stage and lymph node metastasis. Loss-of-function assay indicated that knockdown of has_circ_0008274 suppressed PTC cells proliferation and invasion in vitro. Mechanistically, has_circ_0008274 could inhibit the activation of AMPK/mTOR signaling pathway, which was demonstrated by measuring the expression levels of p-AMPK and p-mTOR. CONCLUSIONS: These results demonstrate that increased has_circ_0008274 expression modulates has_circ_0008274 to enhance PTC cells proliferation and invasion. Has_circ_0008274/ AMPK/mTOR axis may be a novel therapeutic candidate target in PTC treatment.

Chloroquine aggravates the arsenic trioxide (As2O3)-induced apoptosis of acute promyelocytic leukemia NB4 cells via inhibiting lysosomal degradation in vitro.

OBJECTIVE: Acute promyelocytic leukemia (APL) is a distinct subtype of acute myeloid leukemia (AML), as standing out for its distinguished sensitivity to all-trans retinoic acid and arsenic trioxide (ATO, As2O3). The As2O3-mediated degradation of PML-RARA (promyelocytic leukemia-retinoic acid receptor-α) oncoprotein via the proteasome pathway appears to be critical for such distinguished sensitivity. MATERIALS AND METHODS: The present study was to evaluate the influence by chloroquine (CQ), an inhibitor to the release of lysosomal enzymes, on the sensitivity of APL cells to As2O3. APL-derived NB4 cell line was treated with As2O3 or/and CQ in vitro. Then, the cell viability, the induction of apoptosis, and autophagy were examined with MTT assay, with TUNEL staining or with enhanced green fluorescence protein (EGFP)-light Chain 3 (LC3) reporter. The apoptosis- or autophagy-associated proteins were quantified with Western blotting assay. RESULTS: Our results demonstrated that the As2O3 treatment promoted either apoptosis or autophagy in APL NB4 cells and upregulated both apoptosis- and autophagy-associated proteins. However, additional CQ treatment deteriorated the As2O3-induced NB4 cell apoptosis, whereas aggravated the As2O3-induced accumulation of acidic vesicular organelles (AVOs) and blocked the lysosomal degradation in NB4 cells. CONCLUSIONS: Chloroquine aggravates the arsenic trioxide-induced apoptosis of APL NB4 cells via inhibiting lysosomal degradation in vitro. It implies that chloroquine might be adjuvant to sensitize APL cells to arsenic trioxide.

[Prenatal supplementations of iron, iron-containing multimicronutrients and antianemic Chinese patent medicines in women in Shaanxi province, 2010-2013].

Objective: To understand the prevalence of prenatal supplementations of iron, iron-containing multi-micronutrients (IMMN) and antianemic Chinese patent medicines (ACPM) and associated factors in women in Shaanxi province. Methods: A sample of 28 367 childbearing-age women who gave birth during 2010-2013 and had specific information of the prenatal nutrients supplementation were recruited using stratified multistage cluster random sampling in Shaanxi province. The information about their basic characteristics and prenatal supplementation of nutrients were collected by a questionnaire survey. Descriptive analysis method was used to analyze the intake rate of iron, IMMN and ACPM during each period of pregnancy, and logistic regression model was used to identify associated factors. Results: The overall prevalence of prenatal iron, IMMN and ACPM supplementation was low (28.99%), and the intake rate of iron was the lowest (5.33%). The prevalence of prenatal supplementation of iron, IMMN and ACPM were lower before pregnancy and in the first trimester than in the second and third trimester. The intake rates for consecutive 2 periods were very low (all were lower than 2.00%). The intake rates of iron, IMMN and ACPM significantly increased year by year. Women living in central Shaanxi had relatively high intake rates of iron (7.22%) and IMMN (16.55%), and women in southern Shaanxi had relatively high intake rate of ACPM (18.50%). The results of logistic regression analysis showed that higher educational level (OR=1.920, 95%CI: 1.617-2.279), antenatal care times≥6 (OR=1.832, 95%CI: 1.604-2.091), etc. were the positive factors for iron intake, and these positive factors were similar to those for IMMN intake. Additionally, rural residence was the negative factor for IMMN intake (compared with urban residence, OR=0.872, 95%CI: 0.788-0.966). Conversely, higher educational level (OR=0.855, 95%CI: 0.746-0.979), higher household income (OR=0.864, 95%CI: 0.796-0.938) were negative factors for ACPM intake, and rural residence was its positive factor (OR=1.285, 95%CI: 1.141-1.447). Conclusions: The prevalence of prenatal supplementation of iron, IMMN and ACPM were low in women in Shaanxi, especially the intake rate of iron was the lowest. The prophylactic iron supplementation before pregnancy or in the first trimester was rare. Measures should be taken to improve the prenatal supplementation of iron in Shaanxi and to standardize the clinical use of ACPM.

MiR-130b functions as a tumor promoter in glioma via regulation of ERK/MAPK pathway.

OBJECTIVE: To investigate the miR-130b expression in patients with glioma and to analyze its role and underlying molecular mechanism on the carcinogenesis. PATIENTS AND METHODS: The expression levels of miR-130b were detected with quantitative Real-time PCR. The relationship between miR-130b expression and clinicopathologic characteristics were analyzed. MiR-130b inhibitor was transfected into glioma cell lines to investigate its role in HCC. MTT assays were conducted to explore the impact of miR-130b down-expression on the proliferation of human glioma cells. Cell cycle and cell apoptosis assays were performed using flow cytometry. Levels of ERK/MAPK pathway related proteins were evaluated by Western blotting. Data were analyzed using the 2-ΔΔCT method through student's t-test via the GraphPad Prism software (La Jolla, CA, USA). RESULTS: The expression of miR-130b was markedly upregulated in glioma cell lines and tissues, and high miR-130b expression was significantly associated with advanced WHO grade (p = 0.022) and low Karnofsky performance score (p = 0.001). In addition, downregulation of miR-130b inhibited the proliferation of glioma cells and induced cell-cycle arrest and cells apoptosis in vivo. Importantly, ERK/MAPK pathway was found to be inactivated in the glioma cell lines after miR-130b knockout experiment. CONCLUSIONS: The current data indicated that miR-130b may play a critical role in the progression of glioma via ERK/MAPK signaling cascades, suggesting that it may be a useful therapeutic agent in glioma patients.

Tetrandrine induces microRNA differential expression in human hypertrophic scar fibroblasts in vitro.

MicroRNAs (miRNAs) have recently been shown to play a role in normal wound healing process. miRNAs may be linked to pathologic wound healing and closely related to the formation of hypertrophic scars. This study aimed to explore the effects of tetrandrine on the miRNA expression profile in human hypertrophic scar fibroblasts (HSFs) in vitro. HSFs were randomly divided into two groups: the tetrandrine treatment group and the control group. The experimental and control groups were collected and analyzed by miRNA array after a 48-h culture. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to confirm the array results. The targets of differentially expressed miRNA were functionally annotated using bioinformatic approaches. miRNA microarray analysis identified 193 differentially expressed miRNAs and the expression of 186 miRNAs in the experimental group decreased while that of 7 miRNAs increased compared to the control group. The most significantly downregulated miRNA was hsa-miR-1246, and hsa-miR-27b had the highest expression level. Significant differentially expressed miRNAs were predicted to be related to several important signaling pathways related to scar wound healing. The differential miRNA expression identified in this study provides the experimental basis for further understanding the anti-fibrosis effect of tetrandrine.

[Tea, coffee intakes and risk of oral squamous cell carcinoma: a case-control study].

Objective: To evaluate the effects of tea and coffee intakes on oral squamous cell carcinoma (OSCC) stratified by milk intake. Methods: A case-control study involving 593 OSCC patients confirmed by pathological diagnoses and 1 128 gender-age frequency matched controls was conducted in Fujian province during September 2010-March 2016. Unconditional logistic regression was used to calculate adjusted odds ratios (aORs) and corresponding 95% confidence intervals (CIs) to assess the effects of coffee, tea intakes and related variables on OSCC. Additive interaction was estimated by relative excess risk interaction (RERI), attributable proportions interaction (API) and synergy index (SI). Results: Tea intake was significantly associated with decreased risk of OSCC: the adjusted ORs were 0.54 for all subjects (95%CI: 0.41-0.71), 0.47 for milk consumers (95%CI: 0.31-0.71) and 0.57 for non-milk consumers (95%CI: 0.40-0.81). Moreover, starting tea drinking at age ≥25 years, moderate tea concentration and water temperature, drinking green tea and oolong tea showed effects to decrease the risk for OSCC in three groups. Additionally, there was a tendency of a reduced risk with increased daily tea drinking and longer tea-drinking period (all trend P<0.05). No significant association was observed between coffee intake and OSCC. A multiplicative but not additive interactions was found between tea drinking and milk intake. Additionally, we did not observe multiplicative and additive interaction between coffee drinking and milk intake. Conclusion: Tea drinking is a protective factor for OSCC, and there is a multiplicative interaction between tea drinking and milk intake. Therefore, tea drinking and increasing intake of milk can reduce the risk of OSCC at certain extent.

Interleukin-10 gene -592C>A polymorphism and susceptibility to gastric cancer.

Numerous studies have evaluated the association between the human interleukin-10 gene -592C>A polymorphism and gastric cancer risk. However, the results have been inconsistent. This meta-analysis was designed to resolve these controversies. Systematic searches of the electronic databases Embase, PubMed, and Google Scholar were performed to identify relevant studies. A meta-analysis was performed to examine the association between the interleukin-10 gene -592C>A polymorphism and gastric cancer risk. Odds ratios (OR) and its 95% confidence intervals (CI) were used for statistical analysis. Twelve studies were included in the meta-analysis, which included 2116 gastric cancer cases and 4077 controls. No significant association was found between the interleukin-10 gene -592C>A polymorphism and gastric cancer risk in total population analysis. In stratified analysis, a significant association was found in the Asian subgroup (AA vs AC: OR = 0.79, 95%CI = 0.64-0.98; dominant model: OR = 1.26, 95%CI = 1.04-1.57), and no significant association was observed among Caucasians. In addition, the corresponding pooled ORs were not substantially altered after excluding one study that deviated from Hardy-Weinberg equilibrium in the control group. This meta-analysis supports an association between the interleukin-10 gene -592C>A polymorphism and gastric cancer risk in Asians but not in Caucasians. Further large and well-designed studies are needed to confirm these conclusions.

Evidence for gonadotrophin secretory and steroidogenic abnormalities in brothers of women with polycystic ovary syndrome.

STUDY QUESTION: Are there abnormalities in gonadotrophin secretion, adrenal steroidogenesis and/or testicular steroidogenesis in brothers of women with polycystic ovary syndrome (PCOS)? SUMMARY ANSWER: Brothers of women with PCOS have increased gonadotrophin responses to gonadotrophin releasing hormone (GnRH) agonist stimulation and alterations in adrenal and gonadal steroidogenesis. WHAT IS KNOWN ALREADY: PCOS is a complex genetic disease. Male as well as female first-degree relatives have reproductive features of the syndrome. We previously reported that brothers of affected women have elevated circulating dehydroepiandrosterone sulfate levels. STUDY DESIGN, SIZE, DURATION: This was a case-control study performed in 29 non-Hispanic white brothers of 22 women with PCOS and 18 control men. PARTICIPANTS/MATERIALS, SETTING, METHODS: PCOS brothers and control men were of comparable age, weight and ethnicity. Adrenocorticotrophic hormone (ACTH) and GnRH agonist stimulation tests were performed. Gonadotrophin responses to GnRH agonist as well as changes in precursor-product steroid pairs (delta, Δ) across steroidogenic pathways in response to ACTH and GnRH agonist were examined. MAIN RESULTS AND THE ROLE OF CHANCE: Basal total (T) levels did not differ, but dehydroepiandrosterone (DHEA) levels (0.13 ± 0.08 brothers versus 0.22 ± 0.09 controls, nmol/l, P = 0.03) were lower in brothers compared with control men. ACTH-stimulated Δ17-hydroxypregnenolone (17Preg)/Δ17-hydroxyprogesterone (17Prog) (7.8 ± 24.2 brothers versus 18.9 ± 21.3 controls, P = 0.04) and ΔDHEA/Δandrostenedione (AD) (0.10 ± 0.05 brothers versus 0.14 ± 0.08 controls, P = 0.04) were lower in brothers than in the controls. GnRH agonist-stimulated Δ17Prog/ΔAD (0.28 ± 8.47 brothers versus 4.79 ± 10.28 controls, P = 0.003) was decreased and luteinizing hormone (38.6 ± 20.6 brothers versus 26.0 ± 9.8 controls, IU/l, P = 0.02), follicle-stimulating hormone (10.2 ± 7.5 brothers versus 4.8 ± 4.1 controls, IU/l P = 0.002), AD (1.7 ± 1.4 brothers versus 0.9 ± 1.5 controls, nmol/l, P = 0.02) and ΔAD/ΔT (0.16 ± 0.14 brothers versus 0.08 ± 0.12 controls, P = 0.005) responses were increased in brothers compared with controls. LIMITATIONS, REASONS FOR CAUTION: The modest sample size may have limited our ability to observe other possible differences in steroidogenesis between PCOS brothers and control men. WIDER IMPLICATIONS OF THE FINDINGS: Decreased ACTH-stimulated Δ17Preg/Δ17Prog and ΔDHEA/ΔAD responses suggested increased adrenal 3ß-hydroxysteroid dehydrogenase activity in the brothers. Decreased Δ17Prog/ΔAD and increased ΔAD/ΔT responses to GnRH agonist stimulation suggested increased gonadal 17,20-lyase and decreased gonadal 17ß-hydroxysteroid dehydrogenase activity in the brothers. Increased LH and FSH responses to GnRH agonist stimulation suggested neuroendocrine alterations in the regulation of gonadotrophin secretion similar to those in their proband sisters. These changes in PCOS brothers may reflect the impact of PCOS susceptibility genes and/or programming effects of the intrauterine environment. STUDY FUNDING/COMPETING INTERESTS: This research was supported by P50 HD044405 (A.D.), K12 HD055884 (L.C.T.), U54 HD034449 (A.D., R.S.L.) from the National Institute of Child Health and Development. Some hormone assays were performed at the University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core that is supported by U54 HD28934 from the Eunice Kennedy Shriver National Institute of Child Health and Human Development. Partial support for some of the clinical studies was provided by UL1 RR025741 and UL1 TR000150 (Northwestern University Clinical and Translational Sciences Institute) from the National Center for Research Resources, National Institutes of Health, which is now the National Center for Advancing Translational Sciences. The authors have no conflict of interest to declare.

The prognostic value of P-cadherin in non-muscle-invasive bladder cancer.

OBJECTIVES: This research aims to specify the prognostic value of P-cadherin on recurrence and progression in non-muscle-invasive bladder cancers (NMIBC). METHODS: A total of 110 NMIBC cases were collected and P-cadherin protein was assessed by immunohistochemical test in these samples. Correlations between P-cadherin expression and clinicopathologic features were analyzed. For recurrence-free and progression-free survival, Kaplan-Meier log-rank test was used. Then Cox univariate and multivariate analyses were further performed. RESULTS: P-cadherin high expression correlated with tumor progression (P = 0.031). Kaplan-Meier results showed that patients with high P-cadherin expression had worse progression-free survival (P = 0.034) but not recurrence-free survival (P = 0.133) than low-expression patients. Cox regression results showed P-cadherin expression was an independent predictor for progression (P = 0.042) but not recurrence (P = 0.139) in NMIBC. CONCLUSIONS: Our results demonstrated that P-cadherin expression correlated with tumor progression and could be taken as an independent predictor for progression in NMIBC.

Regulation network analysis of testicular seminoma at various stages of progression.

Testicular seminoma has become the most common solid malignancy in young men, especially in the 20s group. We obtained the gene expression profile of human testicular seminoma cells from NCBI, identified the differentially expressed genes of testicular seminoma cells of different stages, and constructed the regulation networks of different stages of testicular seminoma using bioinformatics methodology. Forty differentially expressed genes of testicular seminoma cells of different stages were identified. These genes and pathways are apparently involved in the progression of testicular seminoma.

High expression of P53-induced Ring-h2 protein is associated with poor prognosis in clear cell renal cell carcinoma.

PURPOSE: This study was carried out to examine P53-induced Ring-h2 protein (Pirh2) expression and investigate its clinical and prognostic significance in patients with clear cell renal cell carcinoma (ccRCC). METHODS: Pirh2 mRNA and protein expressions were detected by quantitative reverse-transcription polymerase chain reaction (Q-RT PCR) and Western blotting in 35 frozen renal cancer tissue specimens and 35 adjacent normal renal tissue specimens of the same patients. Pirh2 protein expression was assessed by immunohistochemical analysis in 92 paraffin-embedded specimens of human ccRCC and 30 specimens of adjacent normal renal tissue. Correlations between Pirh2 and clinicopathologic features and prognosis were analyzed statistically. RESULTS: Pirh2 mRNA and protein levels in ccRCC samples were increased significantly as compared with the adjacent normal renal tissues (P < 0.001). Pirh2 mRNA overexpression correlated with high stage and grade of the renal cancer (P < 0.001 and P < 0.001 respectively). Pirh2 protein expression was negative in most normal renal tissue specimens (23/30) but positive in 52.2% (48/92) of ccRCC specimens (P = 0.006). Pirh2 protein expression correlated with tumor grade and stage (P < 0.001 and P < 0.001 respectively). The median follow-up interval was 42.0 months. Overexpression of Pirh2 protein in ccRCC was significantly associated with shorter overall survival and recurrence-free survival (P = 0.001 and P = 0.003, respectively). Multivariate analysis showed that Pirh2 expression was an independent prognostic factor for ccRCC patients (P = 0.037). CONCLUSIONS: Pirh2 was up-regulated in ccRCC at both transcriptional and translational levels compared with normal renal tissues, suggesting that Pirh2 may be a potential prognostic marker for ccRCC.

In situ doxorubicin-CaP shell formation on amphiphilic gelatin-iron oxide core as a multifunctional drug delivery system with improved cytocompatibility, pH-responsive drug release and MR imaging.

An amphiphilic gelatin-iron oxide core/calcium phosphate shell (AGIO@CaP-DOX) nanoparticle was successfully synthesized as an efficient anti-cancer drug delivery system, where doxorubicin (DOX) as a model molecule was encapsulated by electrolytic co-deposition during CaP shell formation. The shell of CaP precipitate played a pivotal role, not only in acting as a drug depot, but also in rendering the drug release rate in a highly pH-dependent controlled manner. Together with MR imaging, highly biocompatible drug-carrying CaP shell and efficient cellular internalization, the AGIO@CaP-DOX nanoparticles developed in this study area promising multifunctional nanodevice for nanotherapeutic approaches.