[Clinical features and prognostic factors of advanced myelodysplastic syndromes in children].

Objective: To investigate the clinical features and prognostic factors of advanced myelodysplastic syndromes (MDS) in children. Methods: Clinical data of children diagnosed with advanced MDS in the Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, between September 2009 and April 2022 were retrospectively collected. Follow-up assessments were performed through telephone interviews and the review of medical records until May 1, 2023. The clinical features of children with advanced MDS were summarized by analyzing chromosomal karyotype tests, second-generation gene sequencing results. Multivariate Cox regression analysis was used to investigate the prognostic factors of advanced MDS in children. Results: A total of 69 children, comprising 49 males and 20 females, aged [M (Q1, Q3)] 8 (5, 10) years, were enrolled in the study. Sixty-seven cases underwent chromosomal karyotype testing, of which 42 cases (62.7%) had abnormal karyotypes, with monosomy 7 the most common in 17 cases (25.4%). Forty-three cases underwent next-generation sequencing, with mutations in the SETBP1, NRAS, PTPN11 and RUNX1 genes more common, identified in 12 cases (27.9%), 9 cases (20.9%), 8 cases(18.6%), and 8 cases(18.6%), respectively. The follow-up time [M (Q1, Q3)] was 26 (13, 56) months and the 5-year overall survival rate was 56%(95%CI: 44.4%-70.5%). The 5-year overall survival rate for children who underwent hematopoietic stem cell transplantation (HSCT) was higher than that of children who did not undergo HSCT (73.9% vs 29.1%, P<0.001). HSCT (HR=0.118, 95%CI: 0.037-0.372, P<0.001) was a protective factor for the overall survival rate of children with advanced MDS. Serum ferritin level>356.3 µg/L (HR=6.497, 95%CI: 2.068-20.415, P=0.001) and moderate to severe splenomegaly (HR=4.075, 95%CI: 1.174-14.141, P=0.027) were risk factors for the overall survival rate of children with advanced MDS. Conclusions: Monosomy 7 was the most common abnormal karyotype and SETBP1 was the gene that had the highest mutation frequency in children with advanced MDS. HSCT, increased ferritin and moderate to severe splenomegaly are prognostic factors influencing the overall survival rate of children with advanced MDS.

[Perioperative efficacy of laparoscopic duodenal-preserving pancreatic head resection].

Objective: To compare the perioperative outcomes of laparoscopic duodenal-preserving pancreatic head resection(LDPPHR) with laparoscopic pancreaticoduodenectomy(LPD) in the treatment of borderline and benign diseases of the pancreatic head. Methods: This is a retrospective cohort study. Perioperative data from 87 patients with non-malignant pancreatic head diseases who underwent LDPPHR or LPD were retrospectively collected in the Department of Biliary-Pancreatic Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from January 2020 to December 2022. There were 49 male and 38 female patients with an age (M(IQR)) of 57.0(16.5) years (range: 20 to 75 years). Forty patients underwent LDPPHR and 47 patients underwent LPD. Quantitative data following a normal distribution were compared using Student's t-test, while quantitative data not following a normal distribution were compared using the Mann-Whitney U test. Comparisons of categorical or ordinal variables were made using χ2 test or Fisher's exact test. Logistic regression analysis was used to estimate the risk factors associated with the rate of complications. Results: There were no statistically significant differences between the LDPPHR group and the LPD group in terms of reoperation rate,total hospital stay duration,postoperative hospital stay duration,90-day mortality rate,30-day and 90-day readmission rates,and 2-year tumor recurrence rate (all P>0.05). The complication rate was higher in the LDPPHR group compared to the LPD group (80.0%(32/40) vs. 51.1%(24/47), χ2=7.89,P=0.005),but there was no difference in the rate of Clavien-Dindo classification of surgical complications ≥Ⅲ between the two groups (10.0%(4/40) vs. 12.8%(6/47), χ2<0.01, P=0.947). Additionally,the rate of delayed gastric emptying (DGE) was higher in the LDPPHR group compared to the LPD group (χ2=10.79,P=0.001),but there was no statistically significant difference in the rate of B,C grade DGE between the two groups (χ2=0.48, P=0.487). There were no statistically significant differences in the rates of postoperative pancreatic fistula,bile leakage,post-pancreatectomy hemorrhage,intra-abdominal infection,and pulmonary infection between the two groups (all P>0.05). The results of the univariate logistic regression analysis showed that LDPPHR (compared to LPD, OR=3.83, 95%CI: 1.46 to 10.04, Z=2.73,P=0.006) and preoperative biliary stent placement (compared to non-use of biliary stent, OR=5.30, 95%CI: 1.13 to 25.00, Z=2.11, P=0.035) were risk factors for the complication rate,but neither was an independent risk factor for complication rate (all P>0.05). Conclusion: The preliminary results suggest that LDPPHR can achieve perioperative safety and effectiveness comparable to LPD.

[Pathological features and immune microenvironment in HER-2 intratumoral heterogeneous breast cancers].

Objective: To observe the clinical pathology features, and immune microenvironment of HER-2 intratumoral heterogeneity breast cancer. Methods: Thirty cases of HER-2 intratumoral heterogeneous breast cancer were retrospectively analyzed in Tianjin Medical University Cancer Institute and Hospital from November 2017 to June 2020. HER-2 expression was detected by immunohistochemistry and verified by dual color silver-enhanced in-situ hybridization (D-SISH). HER-2 intratumoral positive and negative regions were divided. The pathological characteristics, subtype, and the level of tumor infiltrating lymphocytes (TILs) and the expression of programmed cell death-ligand 1 (PD-L1) were evaluated respectively. Results: The proportion of HER-2 positive cells of the breast cancer ranged from 10% to 90%. The pathological type was mainly invasive non-special typecarcinoma. Six cases presented different pathological types between HER-2 positive and negative regions. The HER-2-positive areas included 2 cases of carcinoma with apocrine differentiation, and the negative areas included 2 cases of invasive micropapillary carcinoma, 1 case of invasive papillary carcinoma, and 1 case of carcinoma with apocrine differentiation. In HER-2 positive regions, 17 cases were Luminal B and 13 cases were HER-2 overexpressed types. There were 22 cases of Luminal B and 8 cases of triple negative tumors in the HER-2 negative areas. The levels of TILs in HER-2 positive and negative areas accounted for 53.3% (16/30) and 26.7% (8/30), respectively, with a statistically significant difference (P=0.035). The positive expression of PD-L1 in HER-2 positive area and HER-2 negative area were 6 cases and 9 cases, respectively. Among 8 cases with HER-2 negative regions containing triple negative components, 4 cases were positive for PD-L1 expression. Conclusions: In the case of HER-2 intratumoral heterogeneity, it is necessary to pay attention to both HER-2 positive and negative regions, and evaluate subtype separately as far as possible. For HER-2 intratumoral heterogeneous breast cancer containing triple negative components, the treatment mode can be optimized by refining the intratumoral expression of PD-L1.

Clinical significance of raf-1 kinase inhibitor protein in oral squamous cell carcinoma and its role in cell metastasis.

OBJECTIVE: To detect the expression pattern of raf-1 kinase inhibitor protein (RKIP) in oral squamous cell carcinoma (OSCC) samples and to explore its clinical significance in OSCC metastasis. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot assay were conducted to detect the expression of RKIP in OSCC tissues and cells. The relationship between RKIP expression and OSCC clinicopathological characteristics was statistically analyzed. Transwell assay, wound healing assay, and Western blot were used to detect the influence of RKIP on the metastasis ability of OSCC cells. RESULTS: RKIP was significantly downregulated in OSCC samples. Low expression of RKIP predicted high incidence of metastasis in OSCC patients. In vitro experiments demonstrated that overexpression of RKIP could significantly inhibit invasion and migration abilities of OSCC cells. CONCLUSIONS: RKIP was a novel factor involved in OSCC progression, which was a potential biomarker and therapeutic target for the patients.

[A rare case of cystadenoma in the small intestine].

In recent years, there have been more and more reports about cystadenoma. Cystadenoma can occur in many parts of the body, and cystadenoma in different parts may show different clinical symptoms, however, some patients with cystadenoma have no symptoms. The vast majority of cystadenomas are benign lesions, but a small number of cystadenomas can be malignant. For example, a small number of ovarian cystadenomas and pancreatic cystadenomas may be malignant. This study reported a patient with small intestinal cystadenoma diagnosed by pathology. The patient's physical examination revealed a lesion in the left upper abdomen. He had only abdominal distension and no other discomfort. His laboratory examination results were basically normal, i.e. blood routine, urine routine, stool routine, liver function, kidney function, myocardial enzyme, tumor marker, etc. The patient underwent sectional small intestine resection and the pathological sample was analyzed. The histological findings of the resected intestinal sample were consistent with cystadenoma. Computed tomography scan of the abdomen was performed 4 months after the surgery. No recurrence of the tumor was found. The patient recovered in good condition. By consulting the literature, I found very few reports of small intestinal cystadenoma before, it was very rare. This article described the clinical manifestation, diagnosis and differential diagnosis, treatment and prognosis of a case of small intestinal cystadenoma, it suggested that cystadenoma can occur in the small intestine, other than the ovary, pancreas, liver, lung, thyroid, prostate, seminal vesicle, skin, etc. The cystadenoma in small intestine is easy to be mistaken with other tumors, such as small intestine stromal tumor, small intestine adenocarcinoma, small intestine lipoma, small intestine hemangiomas, etc., and it is difficult to fully confirm through imaging examinations, such as computed tomography and magnetic resonance imaging. Laparotomy and histopathological examination are necessary before definitive diagnosis. This disease can be treated by small bowel resection at the affected region and good prognosis can be achieved.

Clinical efficacy and prognosis of aspirin combined with clopidogrel in patients with cerebral hemorrhage after operation.

OBJECTIVE: The aim of this study was to investigate clinical effect, the quality of life, and prognosis of patients with hypertensive cerebral hemorrhage treated with aspirin combined with clopidogrel after decompressive craniectomy and removal of intracranial hematoma. PATIENTS AND METHODS: The individual patient data of 120 patients with hypertensive cerebral hemorrhage admitted to Affiliated Hospital of Jining Medical University from January 2015 to July 2016 were retrospectively analyzed. The patients were divided into a research group (62 cases) and a control group (58 cases). The control group was treated with aspirin, while the research group was treated with aspirin combined with clopidogrel. The prevalence of adverse reactions was compared between the two groups. Activity of daily living (ADL) was used to evaluate the quality of life. The amount of hematoma before and after operation was compared between the two groups. The prognosis of the two groups and the risk factors of postoperative rebleeding in patients with cerebral hemorrhage were analyzed. RESULTS: The prevalence of adverse reactions in the research group was significantly higher than that in the control group (p<0.05). The ADL scores of both groups 14 days after the operation were higher than those before the operation (p<0.05), and the ADL scores of the research group were significantly lower than those of the control group 14 d after the operation (p<0.05). The amount of hematoma in the two groups after surgery was lower than that before surgery (p<0.05), and the amount of hematoma in the research group was higher than that in the control group (p<0.05). CONCLUSIONS: The combination of aspirin and clopidogrel will increase the prevalence of adverse reactions and reduce the quality of life of patients after decompressive craniectomy and removal of intracranial hematoma in patients with hypertensive intracerebral hemorrhage. Careful medication is required in clinic.

[Clinical features of 54 cases of lung adenocarcinomas with c-ROS oncogene 1 fusion].

Objective: To study the prevalence of c-ros oncogene 1 fusion in lung adenocarcinoma and to evaluate its relationship with clinical characteristics. Methods: We retrospectively analyzed epidermal growth factor receptor (EGFR) mutation, anaplastic lymphoma kinase (ALK) and c-ros oncogene 1 (ROS1) fusion in 1 482 patients with adenocarcinoma from March 2014 to January 2017 in the first affiliated hospital of Zhejiang University. Furthermore, ROS1 fusion positive patients diagnosed between February 2017 and December 2017 were also included in ROS1 positive group. The data of age, sex, smoking history, TNM stage and chest computed tomography were collected by Electronic Medical Record (EMR). The clinical data were compared by the chi-squared test or Mann-Whitney test. Results: Of these 1 482 patients,54 cases were diagnosed with ROS1 rearrangement, including 19 males and 35 females, while 73 cases were diagnosed with ALK rearrangement, including 28 males and 45 females, and 679 cases diagnosed with EGFR mutation including 293 males and 386 females. And there were 676 patients without driven genes mutation. The mean age in ROS1 fusion group (54±12) was lower than EGFR mutation group (60±11, z=-3.982, P<0.001) and WT group (62±10, z=-4.944, P<0.001). Female proportion in ROS1 fusion group (64.8%, 35/54) was higher than WT group (28.4%, 192/676, χ(2)=30.94, P<0.001). Non-smoker percentages in ROS1 fusion group (72.2%, 39/54) was significantly higher than WT group (38.0%,257/676, χ(2)=24.27, P<0.001). ROS1 fusion group was similar to ALK fusion group in sex, age and smoking history, and there were no significant difference in TNM stage among these groups. On chest CT, adenocarcinomas with ROS1 fusion were found to be more peripheral in location (71.4%, 20/28) and solid in density (75%, 21/28), usually with lobulated margins (75.0%, 21/28) and spiculated in contour (57.1%,16/28). Conclusion: In our study lung adenocarcinoma with c-ROS oncogene 1 fusion was a rare subtype lung cancer and was usually detected in young, never smoking, and female patients.

[Analysis on risk factors of positive vertical resection margin after endoscopic treatment of rectal neuroendocrine tumors].

Objective: To analyze the risk factors of positive vertical resection margin of the postoperative specimens after endoscopic treatment of rectal neuroendocrine tumors (NET). Methods: A case-control study was performed. Clinical data of patients with rectal NET (G1) undergoing endoscopic treatment between January 2015 and June 2018 at the Department of Gastroenterology, Beijing Tsinghua Changgung Hospital were retrospectively collected. Inclusion criteria: cases underwent endoscopic mucosal resection (EMR) or endoscopic submucosal dissection (ESD), whose mucosal specimens were pathologically confirmed as NET, and NET was confined to the rectum by nuclide imaging examination before endoscopic treatment. Those with incomplete clinical data or follow-up data were excluded. Resected specimen was fixed and sliced every 2 mm, and when tumor cells were found to infiltrate the vertical cutting edge, the positive vertical margin was defined. Associations of gender, age, resection method, tumor diameter, lesion morphology (nodular lesions, biopsy or post-treatment scar-like changes), mitotic figure, Ki-67 index, etc. and positive vertical margin were analyzed. Univariate analysis was performed using binary logistic analysis and multivariate analysis was performed using logistic regression model. Results: A total of 133 patients with rectal NET were enrolled, including 93 males and 40 females, with an average age of (50.0±10.7) years. Sixty-four patients received EMR treatment and 3 patients (4.7%) had positive vertical margins. While 69 patients received ESD treatment and 13 (18.8%) had positive vertical margins. After endoscopic treatment, 16 cases (12.0%) were vertical positive margin, including 11 males and 5 females with an average age of (52.4±10.4) years. The lesion diameter was (9.0±4.7) mm. Univariate analysis showed that lesion diameter ≥10 mm (χ(2)=5.575, P=0.018) and scar-like changes (χ(2)=3.894, P=0.048) were significantly associated with positive vertical margin. Multivariate analysis showed that the lesion diameter ≥10 mm (OR=10.136, 95%CI: 2.114 to 48.591, P=0.004) was an independent risk factor for positive vertical margin of the specimen after endoscopic treatment of rectal NET. Conclusion: The diameter of rectal NET ≥10 mm indicates a high risk for positive vertical margin after endoscopic treatment.

Osteoinductive effects of tantalum and titanium on bone mesenchymal stromal cells and bone formation in ovariectomized rats.

OBJECTIVE: Although Tantalum (Ta) exhibits better osteoinductivity in healthy subjects when compared with titanium (Ti), the relative effects in osteoporosis remain unknown. MATERIALS AND METHODS: In this study, bone mesenchymal stromal cells of ovariectomized rats (OVX-rBMSCs) were seeded on Ta and Ti substrates for in vitro evaluation of cell viability, reactive oxygen species (ROS) production, alkaline phosphatase (ALP) activity, extracellular mineralization osteogenic gene and protein expression involved in bone morphogenetic protein (BMP2)/small mothers against decapentaplegic homologs 1 (Smad1) pathway. For in vivo assessment, Ta and Ti implants were embedded in femur defects of ovariectomized rats, followed by sequential fluorochrome labeling and histological staining. RESULTS: Compared to Ti, the Ta substrates demonstrated higher viable cell percentages (96.5 ± 0.26 vs. 88.17 ± 2.23%), lower ROS levels (65% vs. Ti), and enhanced ALP activity and extracellular matrix calcification. Reverse Transcription-Polymerase Chain Reaction and Western blot assays validated the better osteoinductive effect of Ta regarding small mothers against decapentaplegic homologs 1 (Smad1), runt-related transcription factor 2, bone morphogenetic protein (BMP2), and ALP expression at both the mRNA (1.5-2-fold) and protein (1.2-1.8-fold) levels. BMP2/Smad1 signaling over-expression or knockdown yielded significantly enhanced or deteriorated OVX-rBMSC osteogenesis on the two surfaces. In addition, the Ta group revealed more new bone formation (1.3-1.5-fold vs. Ti) and slightly better bone-implant contact (31.82 ± 4.07 vs. 25.2-3.84% at 8 weeks post-implantation, p = 0.052) without the contribution of specific surface structures. CONCLUSIONS: In comparison to Ti, Ta reveals better biocompatibility and osteoinductivity to OVX-rBMSCs, and the preferential Ta osteoinductivity may reflect its greater potential to trigger the BMP2/Smad1 cascade. Thus," in front of "Ta". Ta appears preferable to Ti as a bone-implant surface material under osteoporosis conditions.

[Clinicopathologic characterization of malignant mixed tumor of the skin accompanied by eccrine porocarcinoma].

Objective: To investigate the clinicopathologic features, immunophenotype, pathological diagnosis and treatment of malignant mixed tumor (MMT). Methods: Clinical and pathological features including immunohistochemical phenotypes were analyzed in a case of MMT accompanied with eccrine porocarcinoma (EP) involving both hands, diagnosed definitely in January 2018 along with review of relevant literature. Results: A 64-year-old man presented with multiple rash on both hands for 4 years. Three lesions of 0.5 to 2.2 cm were removed for pathological evaluation. The pathological changes on little finger of left and right hands were MMT with EP, whereas that removed from the right ring finger was EP. MMT showed infiltrative growth with vascular wall invasion and consisted of epithelial (glandular or tube differentiation) and mesenchymal components (mucinous and/or cartilage stroma). The endothelial cells showed moderate to severe cytological atypia, nuclear pleomorphism and increased mitotic activity. The glandular component had histological characteristics of syringocarcinoma with moderately atypical chondrocytes but without myoepithelium. EP was composed of basal cells with visible vacuoles in cytoplasm and the presence of tubular and squamous differentiation, along with obvious atypia. Immunohistochemically cavosurface epithelium of glandular differentiation of MMT showed positivity for CK7, EMA and CD117. Myoepithelium showed S-100, CK5/6 and p63 positivity and stromal cells were positive for S-100. Differential diagnoses included metaplastic carcinoma, malignant myoepithelioma and atypical mixed tumor of skin. Conclusions: MMT with EP is extremely rare.The diagnosis of MMT depends on the morphologic features. Immunohistochemical staining is helpful for differential diagnosis. Surgical excision with safety margins is the treatment of choice. Complementary radiotherapy and/or chemotherapy is still controversial. The clinical course of MMT is deemed unpredictable and long-term follow-up is necessary.

Genetic variability of genes involved in DNA repair influence treatment outcome in osteosarcoma.

We conducted a perspective study to investigate the role of ERCC1 (rs11615), ERCC2 (rs13181 and rs1799793), ERCC4 (rs1800067), and ERCC5 (rs17655) in NER pathway in the prognosis of osteosarcoma patients. In total, 146 osteosarcoma patients were recruited between 2008 and 2013. ERCC1 rs11615, ERCC2 rs13181 and rs1799793, ERCC4 rs1800067, and ERCC5 rs17655 gene polymorphisms were assessed by polymerase chain reaction-restriction fragment length polymorphism assay. By multivariate Cox proportional hazards models, we found that carriers of ERCC1 rs11615 TT genotype showed significantly favorable survival compared to wide-type CC genotype, and the adjusted OR (95%CI) was 0.24 (0.08-0.96). Moreover, we found that subjects with ERCC2 rs1799793 AA genotype were associated with decreased hazards of death in multivariate analysis (HR = 0.22, 95%CI = 0.12-0.93). In conclusion, our results suggest that ERCC1 rs11615 and ERCC2 rs1799793 may be useful genetic prognostic markers for osteosarcoma in a Chinese population.

Vascular endothelial growth factor gene is associated with hypertensive cerebellar hemorrhage and rehabilitative treatment.

Early rehabilitative therapy is important for patients with hypertensive cerebral hemorrhage to improve long-term function of the extremities. Vascular endothelial growth factor (VEGF) is closely associated with the pathogenesis of hypertension. To identify the markers contributing to the genetic susceptibility to hypertensive cerebellar hemorrhage (HCH) and rehabilitative treatment, we examined the potential association between HCH and 12 single nucleotide polymorphisms of the VEGF gene. Participants included 244 patients with HCH and 251 healthy controls from our rehabilitation department. The T allelic frequencies of the rs3025020 (intron 6) and rs3025039 (3'-UTR) polymorphisms were significantly higher in the patients with HCH than in the healthy controls (rs3025020 T allele: P = 0.0002, OR = 1.619, 95%CI = 1.256-2.088; rs3025039 T allele: P = 0.001, OR = 1.682, 95%CI = 1.246-2.270). Strong linkage disequilibrium was observed in three blocks (D' > 0.9), and significantly more C-G-C (rs3025020, rs3025030, and rs3025039) haplotypes (P = 0.001) were found in the controls in block 3. Significantly more T-G-C haplotypes were found in the patients with HCH (P = 0.046). Further genotype and clinical phenotype correlation study of the rs3025039 carriers showed that Fugl-Meyer and Barthel index scores were lower in the patients with the TT genotype relative to CT + CC genotypes (P < 0.01). These findings point to a role for VEGF polymorphism in HCH, and may be informative for future investigations on the pathogenesis of rehabilitative treatment.

Diversity and degradation mechanism of an anaerobic bacterial community treating phenolic wastewater with sulfate as an electron acceptor.

Petrochemical wastewater often contains high concentrations of phenol and sulfate that must be properly treated to meet discharge standards. This study acclimated anaerobic-activated sludge to treat saline phenolic wastewater with sulfate reduction and clarified the diversity and degradation mechanism of the microbial community. The active sludge in an upflow anaerobic sludge blanket (UASB) reactor could remove 90 % of phenol and maintain the effluent concentration of SO4 (2-) below 400 mg/L. Cloning and sequencing showed that Clostridium spp. and Desulfotomaculum spp. were major phenol-degrading bacteria. Phenol was probably degraded through the carboxylation pathway and sulfate reduction catalyzed by adenosine-5'-phosphosulfate (APS) reductase and dissimilatory sulfite reductase (DSR). A real-time polymerase chain reaction (RT-PCR) showed that as phenol concentration increased, the quantities of 16S rRNA gene, dsrB, and mcrA in the sludge all decreased. The relative abundance of dsrB dropped to 12.46 %, while that of mcrA increased to 56.18 %. The change in the electron flow ratio suggested that the chemical oxygen demand (COD) was removed mainly by sulfate-reducing bacteria under a phenol concentration of 420 mg/L, whereas it was removed mainly by methanogens above 630 mg/L.

Correlation between carbonic anhydrase IX (CA-9), XII (CA-12) and hypoxia inducible factor-2α (HIF-2α) in breast cancer.

Hypoxia inducible factor 2α (HIF-2α) can trigger the expression of several genes related to many aspects of tumor progress under hypoxia. As an independent factor, the role of HIF-2α is different from other hypoxia-mediated elements, including HIF-1α. Carbonic anhydrase (CA) were also regarded as important enzymes that involve in the tumor microenvironment pH. To find clinical-pathological features of breast cancer in plateau and explore the impact of CAIX, XII (CA-9, CA-12) and HIF-2α on patients with breast cancer. Clinical data were collected and summarized in 94 patients with breast cancer. The expression of HIF-2α, CA-9 and CA-12 were detected, using immunohistochemistry of specimens. The relation between expressions and clinical-pathology was analyzed. Under normoxia, hypoxia, and after being pretreated with the JNK inhibitor SP600125, HIF- 2α, CA-9 and CA-12 expressions were detected, and adhesion and invasion assays were performed in MCF-7 cells and HIF- 2α shRNA cells, respectively. There are higher expressions of HIF-2α in tumor classification 2 and clinical stage 2 (P < 0.05). High expression of CA-12 was observed in clinical stage 2 (P < 0.05). CA-9 expression is significantly correlated with CA-12 expression (r = 0.376, P = 0.0001). HIF-2α expression is not correlated with both CA-9 expression (P = 0.21) and CA-12 expression (P = 0.27). Breast cancer cells in vitro showed that HIF-2α, CA-9 or CA-12 had an increase expression under hypoxia (1% O2). CA-9 or CA-12 expression was observed in HIF-2α shRNA cells. JNK inhibitor SP 600125 reduced the HIF-2α expression and inhibited the adhesion and invasion of breast cancer cell. Slight inhibition effect on CA-9 and CA-12 expression was found. In conclusion, HIF-2α, CA-9 and CA-12 are important hypoxia responsive elements in breast cancer. HIF-2α was involved in metastasis and invasion of breast cancer cells under hypoxia, by the involvement of c-Jun NH2-terminal kinase (JNK) signal pathway. CA-9 and CA-12 may tend to be regulated by HIF-1α more often than by HIF-2α under hypoxia.

YES-associated protein 1 promotes adenocarcinoma growth and metastasis through activation of the receptor tyrosine kinase Axl.

Yes-associated protein 1 (YAP1), a nuclear effector of the Hippo pathway, plays an important role in tumorigenesis and progression of multiple cancers. The present study aimed to investigate the clinical significance of YAP1 and receptor tyrosine kinase Axl expression in human lung adenocarcinomas (LAC). We further explored possible molecular mechanisms mediated by YAP1 in LAC and gastric adenocarcinoma (GAC) cells. Forty-nine cases of human LAC and normal lung tissue (NLT) were collected. The expression of YAP1 and Axl was assessed by immunohistochemical assay through tissue microarray procedure and the clinicopathologic characteristics of all patients were analyzed. Using a loss of function approach, we investigated the effects of small hairpin RNA (shRNA)-mediated knockdown of YAP1 on the expression of Axl, proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-9 (MMP-9), and the proliferative activities and invasive potential in LAC A549 and GAC SGC-7901 cell lines. As a result, the expression of YAP1 and Axl was found in LAC tissues with higher strong reactivity rate compared to the NLT (87.8 percent vs.60.8 percent, P=0.000; 77.6 percent vs 0.0 percent, P=0.000), but they did not associate with the age, gender, tumor size, TNM staging or lymph node metastases of LAC patients (each P>0.05). Spearman rank correlation analysis showed a positive correlation between YAP1 and Axl expression. Furthermore, knockdown of YAP in vitro markedly down-regulated the expression of Axl, PCNA and MMP-9, and inhibited the proliferation and invasion of LAC and GAC cells. Taken together, YAP1 and Axl are highly expressed in LAC compared to the NLT, and knockdown of YAP1 may inhibit the proliferation and invasion of adenocarcinoma cells through downregulation of the Axl pathway, representing a potential therapeutic target for the treatment of cancer.

Overexpression of TNF-alpha and TNFRII in invasive micropapillary carcinoma of the breast: clinicopathological correlations.

AIMS: Angiogenesis is essential for tumour growth and metastasis and tumour necrosis factor (TNF)-alpha is a potent angiogenic factor. Invasive micropapillary carcinoma of the breast (IMPC), a rare subtype of breast cancer, possesses a lymphotropic nature with a high incidence of lymph node metastasis and poor prognosis. The aim was to evaluate the role of TNF-alpha and its receptor TNFRII in the vascular development and metastasis of IMPC. METHODS AND RESULTS: One hundred cases of IMPC and 97 cases of invasive ductal carcinoma, not otherwise specified (IDC) were studied in parallel by immunohistochemistry for TNF-alpha and TNFRII, and microvessel density (MVD) of the tumours was measured. The results showed that the expression of TNF-alpha and TNFRII and the MVD were higher in IMPC than in IDC (P < 0.05). In IMPC, MVD was significantly increased in those with lymph node metastasis compared with those without nodal metastasis (P = 0.001). TNF-alpha expression showed a significant positive correlation with the rate of proliferation, histological grade, lymph node metastasis and MVD (P < 0.05), whereas expression of TNFRII was correlated with TNF-alpha expression and the proliferation of tumour cells in IMPC (P < 0.05). CONCLUSIONS: Expression of TNF-alpha and TNFRII might play an important role in the angiogenesis, tumour cell proliferation and metastasis of IMPC. These markers could represent new targets for therapeutic intervention, i.e. blocking of TNF-alpha and its signal transduction could be a promising tool for treatment.