Comparison of intravesical chemotherapy regimens after radical nephroureterectomy for upper tract urothelial carcinoma and analysis of risk factors for postoperative recurrence.

OBJECTIVE: Upper tract urothelial carcinoma (UTUC) is a relatively rare but aggressive type of urologic cancer that includes renal pelvic tumors and ureteral tumors with a poor prognosis. Full-length nephroureterectomy plus sleeve bladder resection is the standard treatment for the disease, but patients are prone to recurrence of bladder tumors after surgery. Intravesical infusion therapy is the main means to prevent the recurrence and progression of bladder cancer. Epirubicin and gemcitabine are widely used in clinical practice as first-line or salvage therapy for intravesical chemotherapy; however, the efficacy of these agents is rarely discussed. The purpose of this study was to investigate the effects of epirubicin and gemcitabine on the occurrence of bladder cancer after radical nephroureterectomy for UTUC and to analyze the risk factors affecting the recurrence of postoperative bladder cancer. PATIENTS AND METHODS: A total of 215 patients with diagnosed UTUC and treated in our hospital from June 2019 to August 2021 were retrospectively selected as the research subjects, and they were divided into an observation group (120 cases) and a control group (95 cases) according to different treatment methods. The patients in the control group were treated with epirubicin, while those in the observation group received gemcitabine. All patients were followed up by telephone or outpatient examination for 12 months to record the occurrence of adverse reactions. The occurrence of bladder cancer was recorded at 3 months, 6 months, and 12 months after the surgery. According to the occurrence of bladder cancer after surgery, the patients were divided into a bladder cancer group (63 cases) and a non-bladder cancer group (152 cases). Multivariate Logistic regression analysis was used to analyze the risk factors of bladder cancer after surgery. RESULTS: The total incidence of adverse reactions in the control group was 49.47%, which was higher than that in the observation group with 15.00% (p<0.01). The incidence of bladder tumors in the observation group and the control group was 0.00% and 2.11% at 3 months, 5.00% and 8.42% at 6 months, 13.33% and 15.79% at 12 months, without significant difference (p>0.05). After 12 months of perfusion, the levels of acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) in the two groups were significantly lower than those before perfusion (p<0.05). In the observation group, the levels of these three factors were slightly decreased compared with those in the control group, without a significant difference (p>0.05). Between the bladder cancer and non-bladder cancer groups, there were significant differences in tumor location, number of lesions, tumor stage, preoperative ureteral examination, and preoperative history of bladder cancer (p<0.05). The above indexes were all risk factors for postoperative bladder cancer (p<0.05). CONCLUSIONS: Epirubicin and gemcitabine reduced the occurrence of bladder cancer and effectively inhibited tumor angiogenesis after radical nephroureterectomy for UTUC. The tumor location, number of lesions, tumor stage, preoperative ureteral examination, and preoperative history of bladder cancer were risk factors for postoperative bladder cancer.

Systematic evaluation of influencing factors for Chinese rural doctors' job satisfaction and turnover intention: based on the two-factor theory.

OBJECTIVE: In China, rural doctors (RDs) perform crucial health care missions. However, they have received less attention than their colleagues in urban public hospitals. In this specific group, a severe challenge occurs in sync with a high turnover rate and deficient job satisfaction. MATERIALS AND METHODS: This study aims to systematically summarize and evaluate the influencing factors of job satisfaction and turnover intention among Chinese rural doctors. Seven databases, including PubMed, EMBASE, and China National Knowledge Infrastructure (CNKI), were systematically retrieved, and several experts were consulted to acquire holistic publications in this domain. RESULTS: A total of 20 full-text papers and 22,721 samples were included. In addition, 53 influencing factors were evaluated, of which 38 factors may play a significant role. Based on Herzberg's two-factor theory, together with China's cultural tradition and national conditions, we classified these influencing factors into sociodemographic characteristic factors (n=13), incentive factors (n=18), and health care factors (n=22). Meanwhile, we discussed and analyzed the influencing factors of turnover intention and job satisfaction in detail and put forward corresponding measures and suggestions for the government. CONCLUSIONS: We are confident that this research provides a holistic perspective to systematically evaluate the factors influencing the job satisfaction and intention to leave of Chinese rural doctors. Importantly, we hypothesize that the illumination of cases among Chinese rural physicians applies to other countries or regions, which has significant implications for the reformation of the medical system by governments or decision-makers worldwide.

Is chronic obstructive pulmonary disease an independent predictor for adverse outcomes in coronavirus disease 2019 patients?

OBJECTIVE: This study aimed to investigate whether pre-existing chronic obstructive pulmonary disease (COPD) was an independent predictor for adverse outcomes in coronavirus disease 2019 (COVID-19) patients. MATERIALS AND METHODS: We searched electronic databases, including PubMed, Web of Science, EMBASE, and Chinese National Knowledge Infrastructure (CNKI) to screen for eligible articles. A quantitative meta-analysis was performed on the basis of adjusted effect estimates. RESULTS: We observed that COPD was significantly associated with an increased risk of adverse outcomes in COVID-19 patients, which is based on 18 studies with 26,075 cases reporting adjusted effect estimates (pooled effect = 1.53, 95% confidence interval (CI): 1.29-1.8; I2 = 35.4%, random-effects model). CONCLUSIONS: We found that pre-existing COPD was an independent risk factor for predicting the adverse outcomes in COVID-19 patients.

Lipopolysaccharide-stimulated bone marrow mesenchymal stem cells-derived exosomes inhibit H2O2-induced cardiomyocyte inflammation and oxidative stress via regulating miR-181a-5p/ATF2 axis.

OBJECTIVE: Myocardial infarction (MI) is a cardiovascular disease that seriously endangers human health. Exosomes secreted by stem cells have big potential for the treatment of many diseases. The purpose of this study was to study the therapeutic effects of exosomes derived from lipopolysaccharide (LPS)-stimulated bone marrow mesenchymal stem cells (BMSCs) on MI. PATIENTS AND METHODS: The surface markers of BMSCs were detected by Western blot. After BMSCs were stimulated with LPS for 2 days, the exosomes secreted by BMSCs were extracted and observed by transmission electron microscopy (TEM), and their specific surface markers were detected by Western blot. H9c2 cells were co-cultured with exosomes for 24 hours, and then, treated with H2O2 for 4 hours. Next, H9c2 cells were transfected with miRNA-181a-5p mimic (MIM) or negative control (NC). Inflammation and oxidative stress of H9c2 cells were detected by Western blot, cell staining, reactive oxygen species (ROS) quantification, and SOD activity assay. At last, miR-181a-5p expression in BMSCs, exosomes, and H9c2 cells was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). RESULTS: The results revealed that the expression of miR-181a-5p was increased in LPS-stimulated BMSCs (L-BMSC) and in their secreted exosomes. Besides, the expressions of TNF-α and IL-1ß were decreased, while those of SOD1 and SOD2 were increased in H9c2 cells co-cultured with exosomes secreted by LPS-stimulated BMSCs (L-EXO) and transfected with MIM. Moreover, the fluorescence intensity of IL-1ß immunofluorescence was decreased in H9c2 cells co-cultured with L-EXO or transfected with MIM. The level of ROS was also decreased in H9c2 cells co-cultured with L-EXO or transfected with MIM. Furthermore, miR-181a-5p was found to target ATF2 through target gene prediction databases and Western blot and Dual-Luciferase reporter assays. CONCLUSIONS: LPS stimulation can increase the expression of miR-181a-5p in BMSCs, and miR-181a-5p inhibits myocardial inflammation and oxidative stress by targeting ATF2.

D-dimer is associated with the risk of mortality in Coronavirus Disease 2019 patients.

This study aims to investigate the association between D-dimer and the risk of mortality in coronavirus disease 2019 (COVID-19) patients using a meta-analysis. We found that the D-dimer levels in non-survival patients were significantly higher than those in survival patients (SMD = 0.91, 95% CI = 0.79 to 1.03). In conclusion, the elevated D-dimer levels were associated with an increased risk of mortality in COVID-19 patients.

Effect of rutin on cisplatin-induced damage in human mesangial cells via apoptotic pathway.

Cisplatin (CP) is one of the most effective and widely used compounds in the treatment of disease, including cancer, but is known to induce toxicity in patients. Rutin (RUT) is a flavonoid glycoside from Sophora japonica L. that has been shown to possess antioxidative, anti-inflammatory, and antiviral properties. RUT is also known to attenuate cardiotoxicity, isoproterenol-induced cardiac fibrosis, and ischemia/reperfusion-associated hemodynamic alteration, and prevents high glucose-induced renal glomerular endothelial hyperpermeability. In this study, we investigated the effect of RUT on CP-induced nephrotoxicity. CP was used to induce toxicity in human mesangial cells (HMCs), HMCs were pretreated with different concentrations of RUT before being exposed to 10 µg/mL of CP. A positive group was pretreated with antioxidant agent N-acetylcysteine prior to CP administration. At doses between 12.5 and 25 µM, RUT prevented CP-induced reduction in cell viability. Treatment with RUT suppressed intracellular reactive oxygen species and malonic dialdehyde levels and inhibited cell apoptosis. RUT reversed the CP-induced upregulation of p53, cleaved-caspase-3, and increased pro-caspase-3 and pro-caspase-9 levels. In conclusion, the RUT can relieve CP-induced nephrotoxicity by inhibiting the p53/caspase signaling pathway.

KFL2 participates in the development of ulcerative colitis through inhibiting inflammation via regulating cytokines.

OBJECTIVE: Ulcerative colitis (UC) is a kind of chronic inflammatory bowel diseases that seriously endangers human health. The pathogenesis of UC is closely related to the intestinal immune response. Cytokines exert a key role in the regulation of intestinal inflammatory and immune responses. Abnormalities in the function and quantity of various cytokines or imbalance of inflammatory factors and immune factors would lead to UC development. We aimed to investigate whether Kruppel-like transcription factor 2 (KFL2) participates in the development of ulcerative colitis by regulating inflammation, so as to provide a new direction for the clinical treatment. PATIENTS AND METHODS: 40 UC patients were enrolled in this study, including 20 patients with mild ulcerative colitis (MUC) and 20 with severe ulcerative colitis (SUC). 20 normal end of intestinal tissues surgically resected from patients with colorectal cancer in the same period were selected as the control group. Hematoxylin-eosin (HE) staining was used to detect the inflammatory infiltration of intestinal mucosa tissues. Expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α) in peripheral blood mononuclear cells (PBMCs) of each group were detected by qRT-PCR (quantitative Real-Time Polymerase Chain Reaction). Immunohistochemistry was performed to observe the infiltration of IL-6 and TNF-α in intestinal mucosal tissues. Protein and mRNA levels of KLF2 in PBMCs of each group were detected by Western blot and qRT-PCR, respectively. The relationship between the mRNA level of KLF2 in PBMCs and expressions of IL-6, IL-8, IL-10, TNF-α were analyzed using qRT-PCR. RESULTS: Inflammatory cells and cytokines were infiltrated in the intestinal mucosa of UC patients. IL-6, IL-8, IL-10, and TNF-α were overexpressed in PBMCs of UC patients than those of controls. Protein and mRNA levels of KLF2 in PBMCs of UC patients were remarkably lower than those of controls, which were more significant in SUC patients. Meanwhile, KLF2 was closely related to expressions of IL-6, IL-8, IL-10, and TNF-α in PBMCs of UC patients. CONCLUSIONS: KLF2 was downregulated in PBMCs of UC patients than that of normal controls, which participated in the inflammatory response of UC by regulating expressions of IL-6, IL-8, IL-10, and TNF-α. KLF2 may suggest new treatments for ulcerative colitis.

Effects of miR-455 on PIK3R1 gene expression regulation and kidney cancer cell functions.

OBJECTIVE: Migration and proliferation of kidney cancer cells are critical factors affecting effective treatment. Abnormal gene expression exists during cancer pathogenesis. The study showed certain effects of PIK3R1 gene on migration and proliferation of kidney cancer cells, although a few studies have been performed regulatory mechanism of microRNA on PIK3R1 gene. Previous sequencing of PIK3R1 gene found the existence of mir-455 binding site. This study confirmed the regulation of PIK3R1 gene expression in kidney cancer cells of mir-455 by in vitro assay, and analyzed its effects on migration or proliferation of kidney cancer cells. MATERIALS AND METHODS: mir-455 agonist and inhibitor sequences were designed and synthesized to transfect kidney cancer cell line CAKI-1, which were further divided into agonist, inhibitor, and control group. qRT-PCR was used to test expression of mir-455 and PIK3R1 at 12 h, 24 h and 48 h after transfection. PIK3R1 protein expression was measured by Western blot. MTT and cell scratch assay were employed to measure cell proliferation and migration potency. RESULTS: 12 h after transfection with mir-455, no significant difference of the level of PIK3R1was found among the three groups (p>0.05). However, at 24 h and 48 h post-transfection, PIK3R1 expression in agonist group was significantly elevated, along with weakened cell proliferation or migration potency (p<0.05 with significant between-group comparison). By contrast, the level of PIK3R1 was statistically decreased in inhibitor group, in which cell proliferation and migration were enhanced (p<0.05). CONCLUSIONS: In kidney carcinoma cell CAKI-1, mir-455 expression regulation can positively alter PIK3R1 gene expression. Over-expression of PIK3Ra gene could reduce the proliferation or migration potency of CAKI-1 cells.

Idarubicin-intensified haploidentical HSCT with GvHD prophylaxis of ATG and basiliximab provides comparable results to sibling donors in high-risk acute leukemia.

We designed a novel haploidentical hematopoietic stem cell transplantation (haplo-HSCT) system using idarubicin (IDA) intensified conditioning regimens and combination of antithymocyte globulin and basiliximab for GvHD prophylaxis. The outcomes of 110 high-risk acute leukemia patients undergoing haplo-HSCT were compared with 69 contemporaneous high-risk patients receiving HLA-matched sibling transplantation using uniform IDA-intensified regimens. The relapse incidence of haplo-HSCT was 23.4%, and 3-year overall survival (OS) and disease-free survival (DFS) achieved 62.9%, 59.1%, respectively. The cumulative incidences of II-IV and III-IV aGvHD were 28.6 and 14.3%, while limited and extensive cGvHD were 19.4, 13.8%. All these results were equivalent to those of concurrent identical sibling transplantation. Three-year OS and DFS for patients in advance stage reached 48.5, 47.3%. Furthermore, the relapse, 3-year OS of positive minimal residual disease (MRD) patients did not differ from negative MRD patients (18.9% vs 11.5%, 63.6% vs 69.6%), indicating our intensified haplo-HSCT technique could circumvent the dismal prognosis of MRD. These data provide reinforcing evidence that our haplo-HSCT system could dramatically improve the survival of high-risk acute leukemia with low relapse and acceptable transplantation-related mortality, and might be a promising therapeutic option for high-risk patients.

MYCN induces neuroblastoma in primary neural crest cells.

Neuroblastoma (NBL) is an embryonal cancer of the sympathetic nervous system (SNS), which causes 15% of pediatric cancer deaths. High-risk NBL is characterized by N-Myc amplification and segmental chromosomal gains and losses. Owing to limited disease models, the etiology of NBL is largely unknown, including both the cell of origin and the majority of oncogenic drivers. We have established a novel system for studying NBL based on the transformation of neural crest cells (NCCs), the progenitor cells of the SNS, isolated from mouse embryonic day 9.5 trunk neural tube explants. Based on pathology and gene expression analysis, we report the first successful transformation of wild-type NCCs into NBL by enforced expression of N-Myc, to generate phenotypically and molecularly accurate tumors that closely model human MYCN-amplified NBL. Using comparative genomic hybridization, we found that NCC-derived NBL tumors acquired copy number gains and losses that are syntenic to those observed in human MYCN-amplified NBL including 17q gain, 2p gain and loss of 1p36. When p53-compromised NCCs were transformed with N-Myc, we generated primitive neuroectodermal tumors with divergent differentiation including osteosarcoma. These subcutaneous tumors were metastatic to regional lymph nodes, liver and lung. Our novel experimental approach accurately models human NBL and establishes a new system with potential to study early stages of NBL oncogenesis, to functionally assess NBL oncogenic drivers and to characterize NBL metastasis.

Detection of circulating tumour cells may add value in endometrial cancer management.

OBJECTIVE: To evaluate the role of circulating tumour cells (CTCs) in patients with endometrial cancer (EC). STUDY DESIGN: This study included 40 patients with a pre-operative diagnosis of high-risk EC between April 2015 and May 2016. Patients were further divided into high-risk (grade 3, non-endometrioid, myometrial invasion ≥1/2 and stage III-IV) and high-intermediate-risk (grade 2-3, endometrioid, myometrial invasion <1/2 and stage I-II) groups according to postoperative pathological results. CTCs were detected using the CellSearch system, and CTC results were correlated with standard clinicopathological characteristics and serum tumour marker CA125/HE4 status using Chi-squared test, continuity correction or Fisher's exact test. The pharmacodynamic effect was detected after the first cycle of adjuvant therapy. Patients were followed up for 13 months to assess outcomes. RESULTS: Fifteen percent of patients had one or more CTCs. The presence of CTCs was found to be significantly associated with cervical involvement (83.33% vs 11.76%, p=0.00). No significant difference in CTC-positive rates was detected between the high-risk and high-intermediate-risk groups, and no significant correlation was found between CTCs and serum CA125/HE4, either by positive rates or exact serum levels of the conventional tumour markers. No more CTCs were detected after the first cycle of standard chemotherapy in this study, and no distant metastases or recurrence were found in the CTC-positive patients during the follow-up period. CONCLUSION: The presence of CTCs was correlated with cervical involvement. Early-stage EC patients with CTCs may benefit from additional adjuvant therapies. Assessment of CTCs may be useful in the management of high-risk EC patients.

Predictor of massive bleeding following stent placement for malignant oesophageal stricture/fistulae: a multicentre study.

AIM: To evaluate retrospectively the incidence and predictors of massive bleeding after stent placement for malignant oesophageal stricture/fistulae. MATERIALS AND METHODS: This retrospective study comprised 519 patients with malignant oesophageal stricture/fistulae that were successfully treated with stent placement at three hospitals. The patients were divided into two groups based on the occurrence of massive bleeding. Univariate and multivariate analysis was performed to evaluate predictive factors of massive bleeding. RESULTS: Massive bleeding occurred in 54 of 519 patients 1-37 days following stent placement. All of the patients who developed massive bleeding died within 24 hours of the event. Univariate analysis showed massive bleeding was associated with the presence of a concomitant tracheal stent (p<0.001), the existence of concomitant oesophageal fistulae (p<0.001), and prior radiotherapy (p<0.001). Multivariate analysis exhibited that concomitant tracheal stent insertion (odds ratio [OR], 23.134; 95% confidence interval [CI], 9.523-56.199; p<0.001), the presence of oesophageal fistulae (OR, 3.724; 95% CI, 1.677-8.269; p=0.001), and prior radiotherapy (OR, 13.310; 95% CI, 5.464-32.421; p<0.001) were predictors of massive bleeding following stenting. CONCLUSIONS: The presence of oesophageal fistulae, prior radiotherapy, and concomitant tracheal stent are important factors contributing to bleeding after stenting.

Activation of Sonic hedgehog signaling in neural progenitor cells promotes glioma development in the zebrafish optic pathway.

Dysregulation of Sonic hedgehog (Shh) signaling has been implicated in glioma pathogenesis. Yet, the role of this pathway in gliomagenesis remains controversial because of the lack of relevant animal models. Using the cytokeratin 5 promoter, we ectopically expressed a constitutively active zebrafish Smoothened (Smoa1) in neural progenitor cells and analyzed tumorigenic capacity of activated Shh signaling in both transient and stable transgenic fish. Transient transgenic fish overexpressing Smoa1 developed retinal and brain tumors, suggesting smoa1 is oncogenic in the zebrafish central nervous system (CNS). We further established stable transgenic lines that simultaneously developed optic pathway glioma (OPG) and various retinal tumors. In one of these lines, up to 80% of F1 and F2 fish developed tumors within 1 year of age. Microarray analysis of tumor samples showed upregulated expression of genes involved in the cell cycle, cancer signaling and Shh downstream targets ptc1, gli1 and gli2a. Tumors also exhibited specific gene signatures characteristic of radial glia and progenitor cells as transcriptions of radial glia genes cyp19a1b, s100ß, blbp, gfap and the stem/progenitor genes nestin and sox2 were significantly upregulated. Overexpression of GFAP, S100ß, BLBP and Sox2 was confirmed by immunofluorescence. We also detected overexpression of Mdm2 throughout the optic pathway in fish with OPG, therefore implicating the Mdm2-Tp53 pathway in glioma pathogenesis. In conclusion, we demonstrate that activated Shh signaling initiates tumorigenesis in the zebrafish CNS and provide the first OPG model not associated with neurofibromatosis 1.

One dimensional spatial localization of polychromatic stationary wave-packets in normally dispersive media.

In this paper we illustrate how the localization of the stationary two-dimensional solution of the propagation equation strongly depends on the features of its spatio-temporal spectral bandwidth. We especially investigate the role of the ultra-broad temporal support and of the spatial bandwidth of the spectrum on the high localization in one spatial dimension of "Bessel-like" or "blade-like" beams, quasi-stationarily propagating in normally dispersive materials, and potentially interesting for microfabrication applications.

OCT4 promotes tumorigenesis and inhibits apoptosis of cervical cancer cells by miR-125b/BAK1 pathway.

Octamer-binding transcription factor 4 (OCT4) is a key regulatory gene that maintains the pluripotency and self-renewal properties of embryonic stem cells. Although there is emerging evidence that it can function as oncogene in several cancers, the role in mediating cervical cancer remains unexplored. Here we found that OCT4 protein expression showed a pattern of gradual increase from normal cervix to cervical carcinoma in situ and then to invasive cervical cancer. Overexpression of OCT4 in two types of cervical cancer cells promotes the carcinogenesis, and inhibits cancer cell apoptosis. OCT4 induces upregulation of miR-125b through directly binding to the promoter of miR-125b-1 confirmed by chromatin immunoprecipitation analysis. MiRNA-125b overexpression suppressed apoptosis and expression of BAK1 protein. In contrast, miR-125b sponge impaired the anti-apoptotic effect of OCT4, along with the upregulated expression of BAK1. Significantly, Luciferase assay showed that the activity of the wild-type BAK1 3'-untranslated region reporter was suppressed and this suppression was diminished when the miR-125b response element was mutated or deleted. In addition, we observed negative correlation between levels of BAK1 and OCT4, and positive between OCT4 and miR-125b in primary cervical cancers. These findings suggest an undescribed regulatory pathway in cervical cancer, by which OCT4 directly induces expression of miR-125b, which inhibits its direct target BAK1, leading to suppression of cervical cancer cell apoptosis.

Prediction of location of gestational sac for pregnancy of unknown location at first sight during exploratory surgery using the ratio of hCG in haemoperitoneum and venous serum.

OBJECTIVE: To evaluate a practical method to predict the location of gestational sacs for pregnancy of unknown location (PUL) during exploratory surgery. STUDY DESIGN: Sixty-nine cases of PUL with a positive pregnancy test and haemoperitoneum but unknown location of the gestational sac at first sight on exploratory surgery were analysed at the Department of Obstetrics and Gynaecology, Shanghai Jiaotong University. The level of hCG in the haemoperitoneum and venous serum were measured, and the ratio of hCG in haemoperitoneum and venous serum (Rp/v-hCG) was calculated. Rp/v-hCG<1.0 was taken to suggest an intrauterine gestational sac, and Rp/v-hCG>1.0 was taken to suggest an abdominal gestational sac. The sensitivity and specificity of Rp/v-hCG for predicting the location of the gestational sac were evaluated prospectively. RESULTS: Among 69 cases of PUL, 17 cases (17/69) were ultimately diagnosed as abdominal gestational sacs before 9 weeks of gestation, and 52 cases (52/69) were ultimately diagnosed as intrauterine gestational sacs. The diagnostic sensitivity and specificity of Rp/v-hCG at the time of exploratory surgery for predicting the location of the gestational sac were 94.1% and 100%, respectively (kappa=0.96; P=0.039). The rate of missed diagnosis was 5.9%. The location of the gestational sac was determined during the initial exploratory procedure for 15 cases (15/17) with an abdominal gestational sac (1 case of splenic pregnancy was diagnosed during secondary surgery) and 37 cases (37/52) with an intrauterine gestational sac. With the exception of gestational sacs located in the pouch of Douglas (52.9%, 9/17), the gestational sacs (47.1%, 8/17) located in the other places were difficult to find. CONCLUSIONS: Rp/v-hCG should be considered when exploratory surgery reveals no visible gestational sacs at first sight. If Rp/v-hCG is >1.0, more careful pelvic or abdominal exploration is required, rather than dilation and curettage, to locate abdominal gestational sacs.

Hypoxia suppresses E-cadherin and enhances matrix metalloproteinase-2 expression favoring esophageal carcinoma migration and invasion via hypoxia inducible factor-1 alpha activation.

The goal of this study was to explore the molecular mechanism of hypoxia inducible factor-1 alpha (HIF-1 alpha) action on migration and invasion of esophageal carcinoma cells. We used cobalt chloride (CoCl(2) ) to mimic tumor hypoxic microenvironment and analyzed the expressions of E-cadherin, matrix metalloproteinase-2 (MMP-2), and HIF-1 alpha in esophageal carcinoma cells under hypoxia by reverse transcription polymerase chain reaction and Western blotting. To analyze the function of HIF-1 alpha in Eca109 and TE1 cells, we established stable HIF-1 alpha knockdown cells using small interfering RNA. Blocking effect was detected by Western blotting. The concentrations of MMP-2 protein in the conditioned medium were also determined by enzyme-linked immunosorbent assay. Wound-healing and cell invasion assay were used to evaluate the migration and invasion of esophageal carcinoma cells. After exposure to hypoxia, expressions of HIF-1 alpha protein in Eca109 and TE1 cells were upregulated, both mRNA and protein levels of E-cadherin were downregulated, and MMP-2 were upregulated (P < 0.05), whereas HIF-1 alpha mRNA had no significant change (P > 0.05). Small interfering RNA could block HIF-1 alpha effectively under hypoxia, then enhanced E-cadherin expression and inhibited MMP-2 expression, respectively. Furthermore, expression of HIF-1 alpha protein was stable even though MMP-2 repressed by BB2516. Compared with that in normoxia, Snail expression was enhanced when Eca109 or TE1 cells exposed to hypoxia. Once HIF-1 alpha blocked, Snail expressions were inhibited accordingly. Wound recovery and the number of invading cells decreased (P < 0.05) after HIF-1 alpha blocked. The hypoxia suppresses E-cadherin expression and enhances MMP-2 expression favoring esophageal carcinoma migration and invasion via HIF-1 alpha activation. Our observations suggest that HIF-1 alpha inhibition might be an effective strategy to weaken the migration and invasion of esophageal carcinoma cells.

Selective regulation of osteoblastic OPG and RANKL by dehydroepiandrosterone through activation of the estrogen receptor ß-mediated MAPK signaling pathway.

The aim of the work was to investigate the differential regulation by dehydroepiandrosterone (DHEA) of the osteoblastic production via the estrogen receptor beta (ER ß)-mediated signaling pathway. Having developed hMG63-ER ß cells and hMG63-shER ß cells, we analyzed the regulation by DHEA of human osteoblastic viability, the receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), and the differential expression of ER ß, ER α, or p-ERK1/2 (extracellular signal-regulated kinase) in hMG63, hMG63-shER ß, and hMG63-ER ß cells pretreated with or without U0126, flutamide, and ICI 182780, followed by DHEA culture. When the level of ER ß was high, DHEA (10 - 7 mol/l) could effectively amplify the proliferation and inhibit the etoposide-induced apoptosis of hMG63 cells (p<0.01 and p<0.05, respectively), which was blocked by U0126. When the expression of ER ß was silenced, DHEA could not significantly improve the viability of hMG63. In the presence of ER ß, DHEA activated the pERK1/2-MAPK signaling pathway but not p38 and JNK. Besides, the regulation of p-ERK1/2 upon DHEA treatment was mainly modulated by ER ß instead of androgen receptor and ER α. The secretion of OPG was declined following the silence of ER ß (p<0.05). RANKL and ER α, however, were unaffected by culture with or without DHEA and U0126, regardless of the ER ß level. DHEA seems to act selectively on osteoblasts via the dominant ER ß receptor, which mediates amplified cell viability through the MAPK signaling pathway involving pERK1/2 and upregulates the production of OPG rather than RANKL.

Novel cancer suppressor gene for esophageal cancer: manganese superoxide dismutase.

OBJECTIVES: To explore the expression of manganese superoxide dismutase (MnSOD) in esophageal squamous cell carcinoma (ESCC) and its relationship with clinicopathological characteristics and biological behavior. METHODS: On 45 patients with ESCC, immunohistochemistry (SP method), reverse transcription-polymerase chain reaction (RT-PCR), and Western blot were used to detect MnSOD protein and mRNA expression in ESCC and in its adjacent normal tissue 5 cm apart from the edge of cancer lesion and without documented microscopic invasive cancer. Meanwhile, the relationship between the pathological features of esophageal cancer and its biological behavior was analyzed. RESULTS: In ESCC and normal esophageal tissue, MnSOD protein expression was identified 31.1% (14/45) and 86.7% (31/45) (P= 0.000), respectively, with the relative expression levels of MnSOD mRNA 0.310 ± 0.036 and 0.482 ± 0.053 (P= 0.000), relatively. Western blot study showed that the relative expressions of MnSOD protein in cancer lesion and in adjacent normal tissue were 0.384 ± 0.038 and 0.766 ± 0.041, respectively (P= 0.000). With longer lesion, deeper invasion, and poorer differentiation, the expression of MnSOD would get lower, indicating that the levels of MnSOD protein and mRNA expression were closely related to the length of lesion, depth of invasion, and degree of differentiation in ESCC (P < 0.05). Nevertheless, the result showed no association with the presence of lymph node metastasis, cancer location, and macroscopic classification (P > 0.05). CONCLUSIONS: MnSOD protein and mRNA expression both decreased in ESCC, which may be related to carcinogenesis and the development of esophageal cancer. Therefore, detecting the expression of MnSOD in esophageal carcinoma would be of clinical significance in understanding its biological behavior and in guiding therapeutic strategy of esophageal cancer.

Differential regulation of dehydroepiandrosterone and estrogen on bone and uterus in ovariectomized mice.

UNLABELLED: Dehydroepiandrosterone (DHEA) may be useful in the treatment of postmenopausal osteoporosis (PMO). Our present study has found the preferable stimulatory effect of DHEA on bone, in contrast to the proliferative effects of estradiol (E2) on the endometrium and the uterus, which suggests that DHEA has greater potential clinical value than estrogens in prophylaxis and therapeutics for PMO. INTRODUCTION: A series of findings raise the possibility that DHEA may be useful in the treatment of PMO. Our present study thus aimed at the differential effects of DHEA and E2 on bone and the uterus in ovariectomized mice as well as the involvement of aromatase, ERalpha, ERbeta, and AR in the effects. METHODS: Ovariectomized and sham BALB/c mice were given daily treatment with either DHEA or E2 for three months, respectively. Bone mineral density was determined by DEXA after the last treatment. Mice were necropsied in 3 months after the treatment to analyze the ultrastructure of their femur osteoblasts (OBs) with a transmission electron microscope (TEM); DHEA, DHEA sulfate (DHEAS) and E2 levels were assayed by EIA; production in vitro of E2 in the uterus or tibia was assayed to evaluate the profile of P450arom activity; ERalpha and ERbeta mRNA levels in the uterus and tibia were determined by real-time PCR. The primary murine OBs were treated with DHEA and E2, respectively for 72 h. Real-time polymerase chain reaction (PCR) and western blot were carried out to evaluate aromatase, ERalpha, ERbeta and AR expression in OBs. RESULTS: Both DHEA and E2 significantly improved BMD and OB ultrastructure; E2 but not DHEA has significantly increased uterus wet weight, endometrium epithelial and gland thickness. Dehydroepiandrosterone not only increased serum, femoral DHEA, DHEAS and E2 concentration, but also increased uterine DHEA and DHEAS other than E2 concentration in site, while E2 only increased serum, uterine and femoral E2 concentration, but failed to alter the concentrations of DHEA and DHEAS. Moreover, DHEA significantly increased tibia P450arom enzyme activity, while E2 increased uterine and tibia aromatase activity. Furthermore, DHEA increased uterine ERbeta and ERalpha, and ERbeta transcription in the tibia, while E2 increased ERalpha transcription in the uterus and tibia. Dehydroepiandrosterone increased aromatase, ERalpha, ERbeta and AR expression in OBs, and increased significantly, but E2 apparently decreased the ratio of ERbeta/ERalpha. CONCLUSIONS: Although both DHEA and E2 augment BMD, the proliferative effects of E2 on the endometrium and uterus reflect the different modes of action on bone and the uterus, indicating that the preferable stimulatory effect of DHEA on bone appears to the more potential clinical values than estrogens in prophylaxis and therapeutics for PMO. But applicability of the findings from rodents in humans needs further study.