KCNH2A561V Heterozygous Mutation Inhibits KCNH2 Protein Expression via The Activation of UPR Mediated by ATF6.

The potassium channel protein KCNH2 is encoded by KCNH2 gene, and there are more than 300 mutations of KCNH2. Unfolded protein response (UPR) is typically initiated in response to an accumulation of unfolded and/or misfolded proteins in the endoplasmic reticulum (ER). The present study aimed to explore the UPR process and the role of activating transcription factor 6 (ATF6) in the abnormal expression of potassium voltage-gated channel subfamily H member 2 (KCNH2)A561V. The wild-type (wt) KCNH2 and A561V mutant KCNH2 was constructed with his-tag. The 293 cells were used and divided into KCNH2wt+KCNH2A561V, KCNH2wt and KCNH2A561V groups. The expression levels of ATF6 and KCNH2 in different groups were detected by Western blotting, reverse transcription-quantitative PCR, immunofluorescence and immuno-coprecipitation assays. The protein types and abundance of immuno-coprecipitation samples were analyzed by mass spectrometry. The proteomic analysis of the mass spectrometry results was carried out by using the reactome database and GO (Gene Ontology) tool. The mRNA expression levels of KCNH2 and ATF6 in the KCNH2wt+KCNH2A561V group were higher compared with the KCNH2A561V group. However, the full-length protein expression of ATF6 was inhibited, indicating that ATF6 was highly activated and a substantial number of ATF6 was sheared in KCNH2wt+KCNH2A561V group compared with control group. Furthermore, A561V-KCNH2 mutation leading to the accumulation of the immature form of KCNH2 (135 kDa bands) in ER, resulting in the reduction of the ratio of 155 kDa/135 kDa. In addition, the abundance of UPR-related proteins in the KCNH2A561V group was higher compared with the KCNH2wt+KCNH2A561V group. The 'cysteine biosynthetic activity' of GO:0019344 process and the 'positive regulation of cytoplasmic translation activity' of GO:2000767 process in the KCNH2A561V group were higher compared with the KCNH2wt+KCNH2A561V group. Hence, co-expression of wild-type and A561V mutant KCNH2 in 293 cells activated the UPR process, which led to the inhibition of protein translation and synthesis, in turn inhibiting the expression of KCNH2. These results provided a theoretical basis for clinical treatment of Long QT syndrome.

[Diagnostic value of contrast-enhanced ultrasound in hepatic epithelioid hemangioendothelioma].

Objective: To investigate the features of contrast-enhanced ultrasound (CEUS) in hepatic epithelioid hemangioendothelioma (HEHE) in order to improve the preoperative diagnosis rate. Methods: CEUS images of 32 pathologically-proven cases of hepatic epithelioid hemangioendothelioma from January 2004 to August 2021 were collected. Lesions were analyzed to observe the features of enhancement mode, enhancement intensity, and distinct enhancement phases. Results: Among the 32 cases, one had a solitary lesion, 29 had multiple lesions, and two had diffuse-type lesions. Contrast-enhanced ultrasound revealed a total of 42 lesions in 32 cases. In terms of arterial phase enhancement, 18 lesions had overall enhancement, six lesions had uneven dendritic enhancement, 16 lesions had rim-like enhancement, and two lesions had just slight peripheral spot enhancement around the lesions. Among the three cases, there were multiple lesions that had overall enhancement and ring enhancement. In terms of the enhancement phase, 20 lesions showed "fast progression", 20 lesions showed "same progression", and two lesions showed "slow progression". During the late arterial or early portal venous phases with rapid washout, all lesions manifested as hypoechoic. With peaked enhanced intensity, 11 lesions had a lower enhancement intensity than the surrounding normal liver parenchyma; 11 lesions had the same enhancement degree as the surrounding normal liver parenchyma; and 20 lesions had a higher enhancement degree than the surrounding normal liver parenchyma. All 16 ring-enhancing lesions had marked hyperenhancement. In the typical enhancing lesions, four showed hyperenhancement, five showed low enhancement, and nine showed isoenhancement. In the dendrite-enhancing lesions, there were two isoenhancing and four hypoenhancing. Contrast-enhanced ultrasound delineated the boundaries of all lesions more clearly than two-dimensional ultrasound. Conclusion: Contrast-enhanced ultrasound has certain value in the diagnosis of hepatic epithelioid hemangioendothelioma.

[Comprehensive treatment options and influencing factors in elderly patients with breast cancer].

Objective: To investigate the treatment options for breast cancer patients aged 65 and over, and analyze the influencing factors. Methods: The clinical data of 521 elderly patients aged 65 years or older,who underwent surgery in Peking Union Medical College Hospital from January 2009 to December 2015, were collected. They were all female and 65-98 years old. The patients were divided into 65-74 years old group (n=353) and ≥ 75 years old group (n=168). The differences of variables including age, functional status, treatment methods, pathological characteristics, comorbidities and survival time between the two groups were compared, and the differences of comprehensive treatment methods and their impact on clinical efficacy were analyzed. Results: The main operation methods of the two groups were modified radical mastectomy [39.1% (138/353) and 33.9% (57/168), respectively], breast conserving surgery [56.9% (201/353) and 61.3% (103/353), respectively]. Among the patients choosing adjuvant therapy, there was no significant difference between the two groups except chemotherapy (all P>0.05). Univariate analysis showed that the choice of chemotherapy was related to age, surgical methods, pathological types, tumor burden, molecular typing, functional status and comorbidities (all P<0.05). The Eastern Cooperative Oncology Group (ECOG) score and the number of comorbidities were independent factors affecting the choice of chemotherapy for breast cancer in the elderly: [ECOG score: adjusted OR=0.45 (95CI: 0.26-0.75), number of comorbidities: adjusted OR = 0.63 (95CI:0.41-0.98); all P<0.05]. The 5-year disease-free survival rate of 521 elderly patients with breast cancer was 86.3%, 5-year overall survival rate was 88.8%, and the breast cancer specific survival rate was 94.3%. Conclusions: The comprehensive treatment of breast cancer patients aged 65 and above is not affected by age, but is associated with tumor burden, pathological type, molecular typing, comorbidities and ECOG score. Among them, ECOG score and the number of comorbidities are the independent factors influencing the choice of adjuvant chemotherapy.

TRIM56 suppresses the malignant development of hepatocellular carcinoma via targeting RBM24 and inactivating the Wnt signaling.

OBJECTIVE: The aim of this study was to explore the expression pattern of TRIM56 in Hepatocellular carcinoma (HCC) patients and its influence on the prognosis, and to illustrate the molecular mechanisms of TRIM56 in regulating HCC cell behaviors. PATIENTS AND METHODS: TRIM56 levels in HCC specimens and paracancerous specimens were detected. Then, the influences of TRIM56 on clinical data and prognosis in HCC patients were assessed. Next, the regulatory effects of TRIM56 on proliferative potential in Huh7 and Bel-7402 cells were determined, and the role of TRIM56 on the Wnt signaling was examined. Finally, biological characteristics between TRIM56 and RBM24 in HCC development were illustrated by Luciferase assay and rescue experiments. RESULTS: TRIM56 was lowly expressed in HCC tissues and cell lines. HCC patients expressing a low level of TRIM56 suffered advanced T stage and poor survival. Besides, overexpression of TRIM56 inhibited proliferative potential of Huh7 cells, while knockdown of TRIM56 in Bel-7402 yielded the opposite result. TRIM56 was able to negatively regulate key genes in the Wnt signaling. In addition, RBM24 was proven to be the downstream target of TRIM56, which was involved in TRIM56-influenced HCC development. CONCLUSIONS: Downregulated TRIM56 in HCC samples is closely linked to pathological staging and prognosis. TRIM56 alleviates the malignant development of HCC by inactivating the Wnt signaling and targeting RBM24.

Long non-coding RNA PLAC2 suppresses the survival of gastric cancer cells through down-regulating C-Myc.

OBJECTIVE: The aim of this study was to explore the effects of long non-coding ribonucleic acid (lncRNA) placenta-specific protein 2 (PLAC2) on the biological behaviors of gastric cancer (GC) cells by regulating the expression of c-Myc gene and its mechanism. PATIENTS AND METHODS: The expression of PLAC2 in GC tissues and different GC cell lines was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The effects of PLAC2 on apoptosis and cycle, migration, and invasion of GC cells were detected using flow cytometry, wound healing assay, and transwell assay, respectively. After interference in PLAC2 expression, the changes in c-Myc expression were determined through qRT-PCR and Western blotting. RESULTS: The expression level of PLAC2 was downregulated in 38 out of 45 cases of GC tissues compared with that in normal gastric tissues, and it also declined in GC cells. The results of flow cytometry showed that after overexpression of PLAC2, the cell cycle was arrested in the G1/G0 phase, and the apoptosis rate was increased. The results of wound healing assay and transwell assay revealed that both migration and invasion of GC cells were inhibited. After overexpression of PLAC2, the mRNA and protein expression levels of c-Myc declined. CONCLUSIONS: LncRNA PLAC2 affects the biological behaviors of GC cells by regulating the expression of c-Myc gene.

[Polarization of human acute monocytic leukemia THP-1 cells-derived macprophages induced by Nippostrongylus brasiliensis proteins in vitro].

OBJECTIVE: To investigate the polarization of human acute monocytic leukemia THP-1 cells-derived macrophages induced by Nippostrongylus brasiliensis proteins in vitro, so as to provide insights into the elucidation of the mechanisms underlying host immune responses to hookworm infections. METHODS: The in-vitro culture of N. brasiliensis was established and maintained in the laboratory, and the third- (L3) and fifth-stage larvae (L5) were collected under a sterile condition for preparation of L3 and L5 proteins. The in-vitro culture of THP-1 cells was established, stimulated with 500 ng/mL PMA to yield M0 macrophages that were adherent to the plate wall. The LPS + IFN-γ group, IL-4 + IL-13 group, L3 protein group and L5 protein group were given stimulation with 500 ng/mL LPS plus 100 ng/mL IFN-γ, IL-4 and IL-13 (both 100 ng/mL), L3 protein (5 mg/mL) and L5 protein (5 mg/mL), respectively, while the negative control group was given no stimulation. The cell morphology was observed using microscopy, the mRNA expression of M1/M2 macrophages-specific genes was quantified using a quantitative real-time PCR (qPCR) assay, and the surface markers of M1/M2 macrophages were detected using flow cytometry, while the levels of cytokines secreted by M1/M2 macrophages were measured using enzyme-linked immunosorbent assay (ELISA) following stimulations, so as to examine the polarization of THP-1-derived macrophages induced by N. brasiliensis proteins in vitro. RESULTS: Following stimulation with PMA, THP-1 cells appeared wall-adherent M0 macrophages, and polarized to typical M1 macrophages following stimulation with LPS + IFN-γ, and typical M2 macrophages following stimulation with IL-4 + IL-13, IL-3 protein or L5 protein. There was a significant difference in the proportion of M1 macrophages among the negative control group, the LPS + IFN-γ group, the IL-4 + IL-13 group, the L3 protein group and the L5 protein group (χ2 = 3 721.00, P < 0.001), with the highest proportion detected in the LPS + IFN-γ group, and there was also a significant difference in the proportion of M2 macrophages among groups (χ2 = 105.43, P < 0.001). There were significant differences among groups in terms of the mRNA expression of CCL2 (F = 191.95, P < 0.001), TNF-α (F = 129.95, P < 0.001), IL-12b (F = 82.89, P < 0.001), PPARγ (F = 11.30, P < 0.001), IL-10 (F = 9.51, P < 0.001) and Mrc1 genes (F = 12.35, P < 0.001). In addition, there were significant differences in the proportion of positive CD86 and CD206 expression among groups (χ2 = 24 004.33 and 832.50, P < 0.001). Higher IL-1ß and TNF-α levels were measured in the LPS + IFN-γ group than in the IL-4 + IL-13 group, the L3 protein group and the L5 protein group (P < 0.001), and greater TGF-ß1 and IL-10 levels were seen in the IL-4 + IL-13 group, the L3 protein group and the L5 protein group than in the negative control group and the LPS + IFN-γ group (P < 0.05). CONCLUSIONS: Both L3 and L5 proteins of N. brasiliensis may induce the polarization of THP-1-derived macrophages to M2 type in vitro.

Combined mTOR/MEK inhibition prevents proliferation and induces apoptosis in NF2-mutant tumors.

OBJECTIVE: Merlin is encoded by Neurofibromatosis type 2 gene (NF-2), a tumor suppressor gene, which causes some multiple tumors forming disease of the nervous system in case of function loss. Bioinformatics analysis suggested that patients with NF-2 mutation had a worse prognosis, while it was associated with PI3K/mTOR activation, implying abnormal apoptosis in NF-2 mutation related tumors. Hence, we supposed that the inhibitors of PI3K/mTOR pathway might play a role in suppressing the tumor proliferation. MATERIALS AND METHODS: Two representative NF-2 mutation tumor model of NCI-H2452 and HEI193 cell lines were adopted, while two PI3K/mTOR pathway inhibitors Trametinib and Vistusertib were chosen to study the proliferation and apoptosis of the tumor cells. RESULTS: CCK8 cell counting experiment showed that both Trametinib and Vistusertib could inhibit the proliferation of NCI-H2452 cell in vitro, while the combination of Trametinib and Vistusertib was more significant. Flow cytometry results showed that both Trametinib and Vistusertib could enhance apoptosis of NCI-H2452 cell in vitro, while the combination of Trametinib and Vistusertib was more significant. Similar results were also achieved for HEI193 cell lines. In vivo tumorigenicity experiments demonstrated that the tumor volume and weight were significantly decreased by both Trametinib and Vistusertib, while their combination had the most significant effect. Western blot results demonstrated that both Trametinib and Vistusertib could inhibit PI3K/mTOR /MEK pathway and enhance the expression of merlin. CONCLUSIONS: We found that PI3K/mTOR inhibitor could decrease the proliferation of NF-2 mutation tumor cell lines by enhancing apoptosis, while the combination of two drugs might have a better effect.

[Trend on mortalities in all-cause and chronic non-communicable diseases among the labor force population in China, 2007-2016].

Objective: To analyze the trends on mortalities of all-cause and deaths caused by chronic and non-communicable diseases (NCDs) among Chinese labor force population during 2007 to 2016. Methods: Data on cause-of-death that collected from the National Mortality Surveillance System was used to analyze the age and area-related specific crude mortality rates, age-standardized mortality rates and component ratios of NCDs, among the Chinese labor force population, during 2007 to 2016. Trend of crude mortality rates and mortality component ratios of the three major diseases (infectious diseases, maternal and infant diseases, nutritional deficiency diseases; NCDs; injuries) were analyzed. Age-standardized mortality of cancer, COPD, cardiovascular and cerebrovascular diseases were also analyzed by gender. Age-standardized mortality was calculated based on the Year 2010 Population Census of China. Joinpoint regression model was used to obtain annual percentage change and 95%CI was set for assessing the trend. Results: In 2016, the age-standardized all-cause mortality rate was 217.23 per 100 000 among the Chinese labor force population, but decreased by -2.8% (95%CI: -3.8%- -1.7%) annually from 2007 to 2016. The gap between different gender and regions gradually narrowed. The proportion of deaths caused by NCDs increased annually by 0.8% (95%CI: 0.7%-0.9%). The age-standardized mortality rate of NCDs appeared as 171.89/100 000, among the Chinese labor force population in 2016, showing a downward trend by -2.4% (95%CI:-3.3% - -1.4%). However, in females, there appeared the greatest decrease, with an average annual change of -3.3% (95%CI:-4.0% - -2.5%). Diseases as cancer, COPD, cardiovascular and cerebrovascular diseases all showed downward trends in the whole country, with an average range of -2.0% (95%CI: -2.6%--1.3%), -8.0% (95%CI: -8.9% - -7.1%), -1.5% (95%CI: -2.9% - -0.1%), -2.3% (95%CI: -2.8% - -1.8%) in a ten-year period, respectively. Conclusion: All-cause and age-standardized mortality rates caused by NCDs among Chinese labor force population were decreasing during 2007 to 2016. However, the constituent ratios appeared increasing, year by year. Close attention needs to be paid on NCDs which affecting the health of the labor force population in China.

[Impact of the 21-gene recurrence score assay in clinical treatment and prognosis analysis for patients with hormone receptor positive early-stage breast cancer].

Objective: To explore the association between the 21-gene recurrence score (RS) and clinicopathologic characteristics as well as prognosis in patients with axillary lymph node negative, hormone receptor (HR) positive breast cancer. Methods: The clinicopathologic data of 439 early breast cancer patients who underwent 21 gene RS testing was retrospectively analyzed. According to the 21 gene RS, the patients were divided into low risk (295 cases), intermediate risk (111 cases) and high-risk (33 cases) group. The relationship between the 21 gene RS and clinicopathological characteristics, treatment, recurrence and metastasis was analyzed. Univariate and multivariate statistical analyses were used to analyze the risk factors for relapse free survival (RFS). Results: Tumor grade, estrogen receptor (ER), progesterone receptor (PR) and Ki-67 index were significantly different among the 3 risk cohorts (P<0.001 for all). After a median follow-up of 32 months, the recurrence rate in low risk group (3.7%) was significantly lower than that in the intermediate-high risk group (9.0%), the locoregional recurrence (LRR) rate of low, intermediate and high risk group was 2.4%, 6.3% and 9.1%; and the distant metastasis (DM) rate in low risk group was 1.4% and 2.1% in the intermediate-high risk group. Univariate analysis showed RS, ER status and endocrine therapy were prognostic factors for RFS (P<0.05 for all). Multivariate analysis showed that RS was an independent significant predictor for RFS (P=0.04). Conclusions: The 21-gene RS is related to tumor grade, ER, PR and Ki-67 index. RS is an independent risk factor for RFS in patients with hormone receptor positive early-stage breast cancer.

[On National Demonstration Areas: a cluster analysis].

Objective: To understand the 'backward' provinces and the relatively poor work among the construction of National Demonstration Area, so as to promote communication and future visions among different regions. Methods: Methods on Cluster analysis were used to compare the development of National Demonstration Area in different provinces, including the coverage of National Demonstration Area and the scores of non-communicable disease (NCDs) prevention and control work based on a standardized indicating system. Results: According to the results from the construction of National Demonstration Area, all the 29 provinces and the Xinjiang Production and Construction Corps (except Tibet and Qinghai) were classified into 6 categories: Shanghai; Beijing, Zhejiang, Chongqing; Tianjin, Shandong, Guangdong and Xinjiang Production and Construction Corps; Hebei, Fujian, Hubei, Jiangsu, Liaoning, Xinjiang, Hunan and Guangxi; Shanxi, Jilin, Henan, Hainan,Sichuan, Anhui and Jiangxi; Inner Mongolia, Shaanxi, Ningxia, Guizhou, Yunnan, Gansu and Heilongjiang. Based on the scores gathered from this study, 24 items that representing the achievements from the NCDs prevention and control endeavor were classified into 4 categories: Manpower, special day on NCD, information materials development, policy/strategy support, financial support, mass media, enabled environment, community fitness campaign, health promotion for children and teenage, institutional structure and patient self-management; healthy diet, risk factors on NCDs surveillance, tobacco control and community diagnosis; intervention of high-risk groups, identification of high-risk groups, reporting system on cardiovascular and cerebrovascular events, popularization of basic public health service, workplace intervention programs, construction of demonstration units and mortality surveillance; oral hygiene and tumor registration. Contents including oral hygiene, tumor registration, intervention on high-risk groups, identification of high-risk population, reporting system on cardiovascular and cerebrovascular events, popularization of basic public health service, workplace intervention programs, construction of demonstration units and mortality surveillance were discerned as the relatively weak areas in the construction programs of National Demonstration Area. Conclusions: Western regions, especially in some remote provinces had the poorest performance during the construction of National Demonstration Area. Programs regarding chronic disease surveillance, identification and intervention on high-risk groups showed the lowest scores and these outcome-oriented tasks should be further focused on, during the next term of review, in these areas.

FOXC1 overexpression is a marker of poor response to anthracycline-based adjuvant chemotherapy in sporadic triple-negative breast cancer.

PURPOSE: Because of its aggressive characteristics and poor prognosis, triple-negative breast cancer (TNBC) has become a hot topic in cancer research. Chemotherapy is currently the only treatment for patients with TNBC. The transcription factor FOXC1 has been associated with TNBC prognosis, but little is known about its effect on chemosensitivity. The aim of this study was to investigate the effects of FOXC1 on chemosensitivity. METHODS: A case-control study was performed on 25 TNBC patients who experienced relapse and/or metastasis. Another 25 patients without relapse or metastasis were randomly selected as controls. Medical records were reviewed for relevant information, and immunohistochemistry was performed to measure FOXC1 levels. The Kaplan-Meier method and Cox analysis were used to analyze differences in disease-free survival (DFS) and overall survival (OS). The correlation of FOXC1 expression with chemosensitivity was analyzed. Data were analyzed using SPSS 21.0 software, and a P value <0.05 was considered to be statistically significant. RESULTS: In 15 of 22 case patients, FOXC1 was overexpressed, whereas only 8 control patients exhibited FOXC1 overexpression (P < 0.05). FOXC1 expression had no correlation with pathological indicators. An anthracycline-based regimen was administered to 21 study patients and 23 control patients. FOXC1 expression was significantly associated with a worse DFS (HR 2.62, 95% CI 1.05-6.50, P = 0.038) but presented no correlation with OS (HR 2.53, 95% CI 0.76-8.40, P = 0.131) among these 44 patients. CONCLUSIONS: This study shows that FOXC1 is correlated with chemosensitivity to anthracycline and could be used as an indicator of chemosensitivity in sporadic TNBC.

Impact of the 21-Gene Recurrence Score Assay in adjuvant chemotherapy selection for node-negative, hormone receptor-positive breast cancer in the Chinese population.

The 21-Gene Recurrence Score Assay has been intensively studied and recommended by major guidelines for treatment decision in early breast cancer. Its impact in adjuvant chemotherapy selection for Chinese population has not been revealed.The prospective part of this study enrolled consecutive node-negative, hormone receptor-positive patients who underwent 21-gene RS testing at Breast Surgery Department of PUMCH (n=134) between May 2012 and August 2013(21-gene RS group). Risk categories were assigned based on the RS and on standard clinicopathologic criteria according to St. Gallen and Adjuvant! Online. The chemotherapy selection and the oncologists' confidence in decision-making before and after RS testing were recorded. The retrospective part of this study compared the chemotherapy decision in the 21-gene RS group and a control group without RS testing (diagnosed between Jan 2011 and Apr 2012,n=170). A total of 304 patients were included in the analysis (134 21-gene RS group, 170 controls). Based on RS, 97 patients were classified as low risk, 29 patients as intermediate risk, and 8 patients as high risk. Tumor grade (P=0.002), PR expression (P<0.001) and Ki-67 index (P<0.001) were significantly different between the 3 risk cohorts.Comparing the St. Gallen guidelines and RS, there was a 41% concordance between risk groups. By using Adjuvant! Online, the correlation between the predicted BCSM and RS was nominal (r=0.10). A total of 29% 21-gene RS group patients changed their treatment decisions after RS testing (P<0.001, 95% CI, 0.18 to 0.49) with 6% (8/134) patients changing to receive chemotherapy besides endocrine therapy and 23% (31/134) changing to reject chemotherapy. After RS testing, more than one half of the oncologists increased their confidence level in treatment recommendation. In the control group, 67.6% (115/170) patients chose chemotherapy plus endocrine therapy. The chemotherapy percentage was much higher than that of 21-gene RS group (30/134, 22%).This is the first study to demonstrate a reduction in the use of adjuvant chemotherapy in women with node-negative hormone receptor-positive breast cancer, based on use of the RS. The RS had an impact on the physicians' treatment decision-making.

Treatment with UDP-glucose, GDNF, and memantine promotes SVZ and white matter self-repair by endogenous glial progenitor cells in neonatal rats with ischemic PVL.

Periventricular leukomalacia (PVL) is one of the foremost neurological conditions leading to long-term abnormalities in premature infants. Since it is difficult to prevent initiation of this damage in utero, promoting the innate regenerative potential of the brain after birth may provide a more feasible, prospective therapy for PVL. Treatment with UDP-glucose (UDPG), an endogenous agonist of G protein-coupled receptor 17 (GPR17) that may enhance endogenous self-repair potentiality, glial cell line-derived neurotrophic factor (GDNF), a neurotrophic factor associated with the growth and survival of nerve cells, and memantine, a noncompetitive antagonist of N-methyl-d-aspartate (NMDA) receptors that block ischemia-induced glutamate signal transduction, has been reported to achieve functional, neurological improvement in neonatal rats with PVL. The aim of the present study was to further explore whether UDPG, GDNF and/or memantine could promote corresponding self-repair of the subventricular zone (SVZ) and white matter (WM) in neonatal rats with ischemia-induced PVL. SVZ or WM tissue samples and cultured glial progenitor cells derived from a 5 day-old neonatal rat model of PVL were utilized for studying response to UDPG, GDNF and memantine in vivo and in vitro, respectively. Labeling with 5'-bromo-2'-deoxyuridine and immunofluorescent cell lineage markers after hypoxia-ischemia or oxygen-glucose deprivation (OGD) revealed that UDPG, GDNF and memantine each significantly increased glial progenitor cells and preoligodendrocytes (preOLs), as well as more differentiated immature and mature oligodendrocyte (OL), in both the SVZ and WM in vivo or in vitro. SVZ and WM glial cell apoptosis was also significantly reduced by UDPG, GDNF or memantine, both in vivo and in vitro. These results indicated that UDPG, GDNF or memantine may promote endogenous self-repair by stimulating proliferation of glial progenitor cells derived from both the SVZ and WM, activating their differentiation into more mature OLs, and raising the survival rate of these newly generated glial cells in neonatal rats with ischemic PVL.

The IL-6-STAT3 axis mediates a reciprocal crosstalk between cancer-derived mesenchymal stem cells and neutrophils to synergistically prompt gastric cancer progression.

Emerging evidence indicate that mesenchymal stem cells (MSCs) affect tumor progression by reshaping the tumor microenvironment. Neutrophils are essential component of the tumor microenvironment and are critically involved in cancer progression. Whether the phenotype and function of neutrophils is influenced by MSCs is not well understood. Herein, we investigated the interaction between neutrophils and gastric cancer-derived MSCs (GC-MSCs) and explored the biological role of this interaction. We found that GC-MSCs induced the chemotaxis of neutrophils and protected them from spontaneous apoptosis. Neutrophils were activated by the conditioned medium from GC-MSCs with increased expression of IL-8, TNFα, CCL2, and oncostatin M (OSM). GC-MSCs-primed neutrophils augmented the migration of gastric cancer cells in a cell contact-dependent manner but had minimal effect on gastric cancer cell proliferation. In addition, GC-MSCs-primed neutrophils prompted endothelial cells to form tube-like structure in vitro. We demonstrated that GC-MSCs stimulated the activation of STAT3 and ERK1/2 pathways in neutrophils, which was essential for the functions of activated neutrophils. We further revealed that GC-MSCs-derived IL-6 was responsible for the protection and activation of neutrophils. In turn, GC-MSCs-primed neutrophils induced the differentiation of normal MSCs into cancer-associated fibroblasts (CAFs). Collectively, our results suggest that GC-MSCs regulate the chemotaxis, survival, activation, and function of neutrophils in gastric cancer via an IL-6-STAT3-ERK1/2 signaling cascade. The reciprocal interaction between GC-MSCs and neutrophils presents a novel mechanism for the role of MSCs in remodeling cancer niche and provides a potential target for gastric cancer therapy.

SALL4, a novel marker for human gastric carcinogenesis and metastasis.

SALL4, a zinc-finger transcriptional factor for embryonic stem cell self-renewal and pluripotency, has been suggested to be involved in tumorigenesis. The role of SALL4 in human gastric cancer, however, remains largely unknown. In this study, we demonstrated that SALL4 was aberrantly expressed at both mRNA and protein levels in human gastric cancer tissues, and SALL4 level was highly correlated with lymph node metastasis. Enforced expression of SALL4 enhanced the proliferation and migration of human gastric cancer cells, whereas knockdown of SALL4 by siRNA led to the opposite effects. In addition, SALL4 overexpression promoted the growth and metastasis of gastric xenograft tumor in vivo. SALL4 overexpression induced epithelial-mesenchymal transition (EMT) in gastric cancer cells, with increased expression of Twist1, N-cadherin and decreased expression of E-cadherin. Moreover, SALL4 promoted the acquirement of stemness in gastric cancer cells through the induction of Bmi-1 and Lin28B. Taken together, our findings indicate that SALL4 has oncogenic roles in gastric cancer through the modulation of EMT and cell stemness, suggesting SALL4 as a novel target for human gastric cancer diagnosis and therapy.

Recombinant Mycobacterium smegmatis expressing an ESAT6-CFP10 fusion protein induces anti-mycobacterial immune responses and protects against Mycobacterium tuberculosis challenge in mice.

The currently used vaccine against tuberculosis, Bacille Calmette-Guérin (BCG), has variable efficacy, so new vaccine development is crucial. In this study, we evaluated a recombinant vaccine prepared from non-pathogenic Mycobacterium smegmatis (rMS) that expresses a fusion of early secreted antigenic target 6-kDa antigen (ESAT6) and culture filtrate protein 10 (CFP10). C57BL/6 mice were immunized with the rMS expressing the ESAT6-CFP10 fusion protein (rM.S-e6c10) or with BCG. The mice in the rM.S-e6c10 group had a significantly higher titre of anti-ESAT6-CFP10 antibodies than did animals in the BCG or saline groups. Spleen cells from rM.S-e6c10-immunized mice exhibited a cytotoxic response to ESAT6 and CFP10-expressed target cells, but spleen cells from animals in the other groups did not. Levels of IFN-γ and IL-2 production by purified T cells from spleens were significantly higher in rM.S-e6c10 group than in BCG group. Finally, after M. tuberculosis (MTB)-challenged mice, dramatic reduction in the numbers of MTB colony-forming units (CFUs) in the lungs was observed for the mice immunized with the rMS. The protective efficacy of rM.S-e6c10 and BCG vaccination was similar based on measures of MTB burden and lung pathology. Our data indicate that the recombinant M. smegmatis vaccine expressing the ESAT6-CFP10 fusion protein has potential in clinic application.

Percutaneous anterior transarticular screw fixation for atlantoaxial instability: a case series.

We reviewed the outcome of a retrospective case series of eight patients with atlantoaxial instability who had been treated by percutaneous anterior transarticular screw fixation and grafting under image-intensifier guidance between December 2005 and June 2008. The mean follow-up was 19 months (8 to 27). All eight patients had a solid C1-2 fusion. There were no breakages or displacement of screws. All the patients with pre-operative neck pain had immediate relief from their symptoms or considerable improvement. There were no major complications. Our preliminary clinical results suggest that percutaneous anterior transarticulation screw fixation is technically feasible, safe, useful and minimally invasive when using the appropriate instruments allied to intra-operative image intensification, and by selecting the correct puncture point, angle and depth of insertion.

Tcf4 can specifically recognize beta-catenin using alternative conformations.

Accumulation of the Wnt pathway effector beta-catenin is a hallmark of a number of cancers, including colon cancer. As beta-catenin accumulates in the cell, it forms a complex with Tcf family transcription factors and activates the transcription of several critical genes involved in cell proliferation. Because Tcf4 is the predominant Tcf factor present in colon cancer cells, drugs that specifically disrupt the beta-catenin-Tcf4 complex could be useful in treating colon cancers. Earlier structural and biochemical studies demonstrated that the central region of the beta-catenin binding domain of Tcf is essential for anchoring Tcf to beta-catenin via two conserved lysines in beta-catenin (called the charged 'buttons'). Here we report the crystal structure of a beta-catenin-Tcf4 complex at 2.0 A resolution. Our structural and mutagenesis studies show that Tcf4 docks specifically to beta-catenin using several distinct conformations in its essential central region. These conformations allow different glutamate residues in the central region of Tcf4 to form a salt bridge with the same critical charged button, Lys 312 of beta-catenin. We propose that this interaction may be the first event in beta-catenin-Tcf4 recognition.

Prostate boundary segmentation from 2D ultrasound images.

Outlining, or segmenting, the prostate is a very important task in the assignment of appropriate therapy and dose for cancer treatment; however, manual outlining is tedious and time-consuming. In this paper, an algorithm is described for semiautomatic segmentation of the prostate from 2D ultrasound images. The algorithm uses model-based initialization and the efficient discrete dynamic contour. Initialization requires the user to select only four points from which the outline of the prostate is estimated using cubic interpolation functions and shape information. The estimated contour is then deformed automatically to better fit the image. The algorithm can easily segment a wide range of prostate images, and contour editing tools are included to handle more difficult cases. The performance of the algorithm with a single user was compared to manual outlining by a single expert observer. The average distance between semiautomatically and manually outlined boundaries was found to be less than 5 pixels (0.63 mm), and the accuracy and sensitivity to area measurements were both over 90%.