[The risk factors for regional lymph node metastasis of mismatch repair deficient colorectal cancer].

Objective: To explore the risk factors for regional lymph node (RLN) metastasis in colorectal cancer patients with mismatch repair deficiency (dMMR). Methods: The data of 357 dMMR colorectal cancer patients who underwent surgery in National Cancer Center from January 2012 to December 2016 was retrospectively analyzed. Univariate and multivariate analysis were used to identify the risk factors for RLN metastasis. Results: Among the 357 patients, 204 were male and 153 were female, 61.6% (220/357) lesion located in right half colon, while the other 16.2% (58/357) located in rectum. Univariate analysis showed that tumor size, differentiation, lymphovascular invasion, tumor deposit, postoperative pathologic T stage (pT), the number of negative lymph nodes and the expression of the MSH6 protein were significantly associated with RLN metastasis (P<0.05). All of the patients with well differentiation tumors (15 patients) or staged pT1 (13 patients) had no RLN metastasis. Multivariate analysis showed that tumor differentiation (OR=2.582, 95%CI=1.567-4.274, P<0.001), pT (OR=3.778, 95%CI=1.448-12.960, P=0.015) and the expression of MSH6 protein (OR=2.188, 95%CI=1.159-4.401, P=0.021) were independent risk factors for RLN metastasis. Conclusions: The postoperative pT stage, tumor differentiation and the expression of MSH6 protein are independent risk factors for RLN metastasis of dMMR colorectal cancer. Preoperative assessment of these factors may further improve the accuracy of predicting the risk of RLN metastasis.

[Long-term conditional disease-free survival for rectal cancer patients underwent neoadjuvant chemoradiotherapy].

Objective: To explore the application value of the conditional disease-free survival (cDFS) analysis in predicting prognosis of stage-specific rectal cancer patients underwent neoadjuvant chemoradiotherapy (nCRT). Methods: Clinicopathologic data of 436 patients with rectal cancer received nCRT and radical operation in Cancer Hospital, Chinese Academy of Medical Sciences between January 2004 and December 2016 were retrospectively reviewed. With reference to conditional probability, the 3-year cDFS of patients at different ypTNM stage after completion of nCRT was estimated using the Kaplan-Meier method. Results: There were 66 patients of ypTNM stage 0 (pathological complete response), 87 patients of ypTNM stage Ⅰ, 135 patients of ypTNM stage Ⅱ and 148 patients of ypTNM stage Ⅲ. The 3-year accumulated DFS of patients with ypTNM stage 0, ypTNM stage Ⅰ, ypTNM stage Ⅱ, and ypTNM stage Ⅲ were 97.0%, 93.1%, 85.2%, and 64.2%, respectively. On the condition of postoperactive disease-free survival for 1 year, 2 years, 3 years, 4 years, and 5 years, the corresponding 3-year cDFS of patients at ypTNM stage 0 were 97.0%, 95.5%, 96.9%, 98.4%, 100.0%, respectively. The corresponding 3-year cDFS of patients at ypTNM Ⅲ were 68.2%, 79.3%, 86.3%, 92.1%, 96.4%, respectively. The more advanced ypTNM staging resulted in the more improvement of 3-year cDFS being acquired. Conclusion: cDFS is a better method to reflect the dynamic changes of the prognosis of rectal cancer patients with nCRT in different ypTNM stage, and it is useful to guide the clinicians to assess the prognosis and propose appropriate surveillance.

[Clinicopathological and prognostic features of young onset patients with middle-low rectal cancer received neoadjuvant chemoradiotherapy].

Objective: To explore the clinicopathological and prognostic features of young onset patients with middle-low rectal cancer who received neoadjuvant chemoradiotherapy (NCRT). Methods: After NCRT, a total of 441 patients with primary middle-low rectal cancer treated with radical surgery at the Cancer Hospital, Chinese Academy of Medical Sciences (CHCAMS) from January 2004 to December 2016 were included. According to the age of disease onset, the patients were divided into the young group (51cases) and the middle-old group (390 cases), and the clinicopathological characteristics and survival of these patients were analyzed. Results: In the young group, 68.6% of patients received radical surgery within 7 weeks after NCRT, which was higher than 52.8% in the middle-old group (P=0.047). The stage ypTNM Ⅲ in the young group was 51.0%, higher than 34.1% in the middle-old group (P=0.027). The stage ypN+ in the young group was 51.0%, higher than 34.1% in the middle-old group (P=0.047), The incidence of disease progression in the young group was 39.2%, higher than 25.1% in the middle-old group (P=0.049). The incidence of distant metastasis in the young group was 35.3%, higher than 21.5% in the middle-old group(P=0.044). Most cases of disease progression occurred in the first 3 years after surgery for the young group, especially in the second year after surgery, the incidence of disease progression in the young group was 55.0%, higher than 26.5% in middle-old group (P=0.025). The 3-year and 5-year disease-free survival (DFS) rates for the young group were 63.7% and 58.2%, lower than 81.0% and 74.3% in the middle-old group (P=0.016), respectively. The 3-year and 5-year overall survival in the middle-old group (OS) rates for the young group were 85.4% and 69.2%, lower than 93.6% and 84.1% in the middle-old group (P=0.033), respectively. The multivariate analysis showed that, response of primary tumor (HR=4.804, 95% CI: 1.360-16.973) and total number of dissected lymph nodes (HR=4.336, 95% CI: 1.739-10.809) in the young group were independent prognostic factors related to DFS. The total dissected number of lymph nodes(HR=3.295, 95% CI: 1.076-10.091)was an independent prognostic factor related to OS. In the middle-old group, response of primary tumor (HR=2.626, 95% CI: 1.354-5.091), ypTNM stage (ypTNM Ⅲ: HR=5.837, 95% CI: 2.968-11.479) and tumor location distance from the anal verge (HR=0.500, 95% CI: 0.308-0.812) were independent prognostic factors related to DFS. Lymphovascular invasion (HR=0.500, 95% CI: 0.308-0.812) and ypTNM stage (ypTNM Ⅲ: HR=16.322, 95% CI: 5.049-52.771) were independent prognostic factors related to OS. Conclusions: Young onset rectal cancer patients are associated with shorter operation time interval, advanced pathological stage and poorer prognosis. More intensive adjuvant treatment and post-treatment surveillance should be conducted to young onset rectal cancer with NCRT.

[Clinicopathological characteristics and prognostic analysis of patients with pathological complete response and near complete response after neoadjuvant treatment of rectal cancer].

Objective: To investigate the clinicopathological characteristics and prognosis of rectal cancer patients with pathological complete response and near complete response after neoadjuvant therapy. Methods: The clinicopathological data of patients who underwent neoadjuvant chemoradiotherapy plus radical surgery of rectal cancer in the Cancer Hospital of Chinese Academy of Medical Sciences from January 2004 to December 2016 were retrospectively collected. The clinicopathological characteristics and prognostic factor of patients with pathological complete response and near complete response were analyzed. Results: The clinical data of 142 patients were collected. There were 93 males and 49 females, aged from 24 to 81 years. The median disease-free survival was 53.9 months and the median overall survival was 55.0 months. Univariate analysis showed that the maximum diameter of scar or lesion, the status of lymph node metastasis and the distance between the lower edge of tumor and anal edge were associated with disease-free survival time; the maximum diameter of scar or lesion and the status of lymph node metastasis were associated with overall survival time. Multivariate Cox proportional hazards regression analysis showed that patients with scar or lesion diameter>3 cm (HR=4.406,95%CI:1.619-12.006), positive lymph node metastasis status (HR=4.102,95%CI:1.461-11.513) and tumor lower margin to anal margin distance ≤4 cm (HR=18.171,95%CI:2.357-140.073) had shorter disease-free survival time.The patients with scar or lesion diameter>3 cm (HR=8.573,95%CI:1.630-45.099) and lymph node metastasis status (HR=4.721, 95%CI:1.068-20.860) had shorter overall survival time. Conclusions: The overall prognosis of patients with pathological complete response or near complete response after neoadjuvant therapy for rectal cancer is better. The distance between the lower margin of the tumor and the anal edge, the status of lymph node metastasis and the maximum diameter of scars or lesion were the related factors affecting the prognosis of patients with rectal cancer.

[The safety and efficacy of radical surgery after modified total neoadjuvant therapy for locally advanced rectal cancer].

Objective: To analyze the clinical-pathological data of patients with locally advanced rectal cancer who underwent modified total neoadjuvant therapy (TNT), and to evaluate the safety and efficacy of radical surgery after modified total neoadjuvant therapy. Methods: The clinical-pathological data of 30 locally advanced rectal cancer patients who underwent modified TNT (mTNT) followed by radical resection were retrospectively analyzed. The surgical procedure, postoperative complications, tumor regression grade, tumor downstaging and prognosis were analyzed. Results: The 30 patients included 24 males and 6 females with a median age of 55.5 years. All patients underwent radical surgery after neoadjuvant therapy, 14 patients received low anterior resection, 14 patients received abdominal perineal resection, and the other 2 patients received Hartmann procedure. All patients achieved R0 resection with a median operative time 220 minutes and the median intraoperative blood loss was 200 ml. The morbidity of postoperative complications was 20% (6/30), including dysuria in 2 patients, delayed healing of perineal incision in 2 patients, intestinal obstruction in 1 patient and pelvic hemorrhage in 1 patient. The median time to first flatus after surgery was 3 days and the median postoperative hospital stay was 8 days. Postoperative pathological results showed that 15 patients (50.0%) had severe tumor regression, including 4 patients (13.3%) achieved pathological complete response (pCR), 12 patients (40.0%) had moderate tumor regression, and 3 patients (10.0%) had minor tumor regression. Twenty patients had detailed pre-treatment clinical stage, and among those 20 patients, 15 patients (75.0%) and 13 patients (65.0%) achieved downstaging of tumor T stage and N stage, respectively. Only 2 patients appeared distant metastasis, and no patient had local recurrence. Conclusions: For locally advanced rectal cancer patients, mTNT doesn't increase the morbidity of postoperative complication and is a safe and effective treatment strategy with satisfactory short-term result.

Risk factors for incidence and case-fatality rates of healthcare-associated infections: a 20-year follow-up of a hospital-based cohort.

Information is lacking on the integrated evaluation of mortality rates in healthcare-associated infections (HAIs). Our aim was to differentiate the risk factors responsible for the incidence from those for the case-fatality rates in association with HAIs. We therefore examined the time trends of both incidence and case-fatality rates over a 20-year period at a tertiary-care teaching medical centre in Taiwan and the mortality rate was expressed as the product of the incidence rate and the case-fatality rate. During the study period the overall mortality rate fell from 0·46 to 0·32 deaths/1000 patient-days and the incidence rate fell from 3·41 to 2·31/1000 patient-days, but the case-fatality rate increased marginally from 13·5% to 14·0%. The independent risk factors associated with incidence of HAIs were age, gender, infection site, admission type, and department of hospitalization. Significant prognostic factors for HAI case-fatality were age, infection site, intensive care, and clinical department. We conclude that the decreasing trend for the HAI mortality rate was accompanied by a significant decline in the incidence rate and this was offset by a slightly increasing trend in the case-fatality rate. This deconstruction approach could provide further insights into the underlying complex causes of mortality for HAIs.

Effect of silencing HIF-1α on proliferation, invasion and migration of glioblastoma U87 cells.

The objective of the study was to evaluate the effect of silencing hypoxia-inducible factor 1α (HIF-1α) on proliferation, invasion, and migration of glioblastoma U87 cells. HIF-1α-shRNA lentiviral vector was designed for liposome-mediated transfection into U87 cells. The efficiency of interference was assessed by RT-PCR and Western blot. Cell proliferation was measured by MTT assay. Cell migration was observed by the migration test. The capabilities of invasion and migration were detected using the Transwell model. The research involved experiments in the interference group (shRNA transfected), the control interference group (empty vector transfected), and the untreated (non-transfected) group. Compared with the control interference group and the untreated group, the expressions of HIF-1α mRNA and protein were significantly down-regulated, and the proliferation and invasion of U87 cells were significantly inhibited in the interference group. HIF-1α mRNA and protein are effectively suppressed by HIF-1α-shRNA in U87 cells, which appears to inhibit proliferation, invasion, and migration of U87 cells.

CD22 associates with protein tyrosine phosphatase 1C, Syk, and phospholipase C-gamma(1) upon B cell activation.

Cross-linking B cell antigen receptor (BCR) elicits early signal transduction events, including activation of protein tyrosine kinases, phosphorylation of receptor components, activation of phospholipase C-gamma (PLC-gamma), and increases in intracellular free Ca2+. In this article, we report that cross-linking the BCR led to a rapid translocation of cytosolic protein tyrosine phosphatase (PTP) 1C to the particulate fraction, where it became associated with a 140-150-kD tyrosyl-phosphorylated protein. Western blotting analysis identified this 140-150-kD protein to be CD22. The association of PTP-1C with CD22 was mediated by the NH2-terminal Src homology 2 (SH2) domain of PTP-1C. Complexes of either CD22/PTP-1C/Syk/PLC-gamma(1) could be isolated from B cells stimulated by BCR engagement or a mixture of hydrogen peroxidase and sodium orthovanadate, respectively. The binding of PLC-gamma(1) and Syk to tyrosyl-phosphorylated CD22 was mediated by the NH2-terminal SH2 domain of PLC-gamma(1) and the COOH-terminal SH2 domain of Syk, respectively. These observations suggest that tyrosyl-phosphorylated CD22 may downmodulate the activity of this complex by dephosphorylation of CD22, Syk, and/or PLC-gamma(1). Transient expression of CD22 and a null mutant of PTP-1C (PTP-1CM) in COS cells resulted in an increase in tyrosyl phosphorylation of CD22 and its interaction with PTP-1CM. By contrast, CD22 was not tyrosyl phosphorylated or associated with PTP-1CM in the presence of wild-type PTP-1C. These results suggest that tyrosyl-phosphorylated CD22 may be a substrate for PTP-1C regulates tyrosyl phosphorylation of CD22.

Comparative proteome analysis of two antagonist Bacillus subtilis strains.

Natural wild-type strains of Bacillus subtilis are extensively used in agriculture as biocontrol agents for plants. This study examined two antagonist B. subtilis strains, KB-1111 and KB-1122, and the results illustrated that KB-1122 was a more potent inhibitor of the indicator pathogen than KB- 1111. Thus, to investigate the intrinsic differences between the two antagonist strains under normal culture conditions, samples of KB-1111 and KB-1122 were analyzed using MALDI-TOF-MS. The main differences were related to 20 abundant intracellular and 17 extracellular proteins. When searching the NCBI database, a number of the differentially expressed proteins were identified, including 11 cellular proteins and 10 secretory proteins. Among these proteins, class III stress-response-related ATPase, aconitate hydratase, alpha-amylase precursor, and a secretory protein, endo-1, 4-beta-glucanase, were differentially expressed by the two strains. These results are useful to comprehend the intrinsic differences between the antagonism of KB-1111 and KB-1122.

Proenkephalin A gene products activate a new family of sensory neuron--specific GPCRs.

Several peptide fragments are produced by proteolytic cleavage of the opioid peptide precursor proenkephalin A, and among these are a number of enkephalin fragments, in particular bovine adrenal medulla peptide 22 (BAM22). These peptide products have been implicated in diverse biological functions, including analgesia. We have cloned a newly identified family of 'orphan' G protein--coupled receptors (GPCRs) and demonstrate that BAM22 and a number of its fragments bind to and activate these receptors with nanomolar affinities. This family of GPCRs is uniquely localized in the human and rat small sensory neuron, and we called this family the sensory neuron--specific G protein--coupled receptors (SNSRs). Receptors of the SNSR family are distinct from the traditional opioid receptors in their insensitivity to the classical opioid antagonist naloxone and poor activation by opioid ligands. The unique localization of SNSRs and their activation by proenkephalin A peptide fragments indicate a possible function for SNSRs in sensory neuron regulation and in the modulation of nociception.

Whole-body positron emission tomography with 18F-fluorodeoxyglucose is an effective method to detect extra-pelvic recurrence in uterine sarcomas.

PURPOSE OF INVESTIGATION: To assess the clinical use of F-18-fluorodeoxyglucose positron emission tomography (FDG-PET) in the post-therapy surveillance of uterine sarcoma. METHODS: Eight whole-body FDG-PET studies were performed in seven women with previously treated uterine sarcoma. Conventional image studies (computed tomography) and physical examinations were performed for follow-up. All FDG-PET studies were indicated to localize suspected recurrences noted by conventional methods. RESULTS: The per case sensitivity of the FDG-PET studies and CT scans was 85.7% (6/7) and 100% (7/7), respectively (p = 0.174). FDG-PET was able to detect seven extrapelvic metastastic sites below the diaphragm (7/7, sensitivity: 100%), including the liver, spleen, paraaortic lymph node, spine and paracolic gutter, as well as pulmonary lesions in five patients, while the CT scan detected only three lesions (3/7, sensitivity: 42.9%; p = 0.070). FDG-PET detected only four recurrent pelvic lesions (4/6) and CT scan detected six (6/6) recurrent pelvic lesions (66.7% vs 100%, p = 0.455). CONCLUSIONS: The FDG-PET showed a better detection rate than the abdominal CT scan for extrapelvic metastatic lesions and a similar detection rate as well as abdominal CT scan. FDG-PET can serve as a useful detection tool for patients with uterine sarcomas because nearly 80% of recurrence involve an extrapelvic site.

[Multicenter clinical trial of acute lymphoblastic leukemia in elder children and adolescents].

Objective: To analyze the clinical characteristics and long-term outcomes with multicenter study for acute lymphoblastic leukemia (ALL) in children over 10 years old and adolescents. Method: Newly diagnosed ALL patients aged from 10 to 18 years old in three hospitals were included in the study from May 1(st) 2005 to April 30(th) 2015. They were received ALL-2005/2009 protocol following up to December 31(st) 2016. The clinical characteristics, outcomes and the prognostic analysis were evaluated between the two protocols. Results: Totally, 237 patients were involved in the study, 76 cases for ALL-2005 and 161 cases for ALL-2009 protocol. Complete remission (CR) after induction therapy was 94.5%. 64 (28.6%) patients relapsed with a median time of 14.5 months and 70 (29.5%) patients passed away during the following time. In long-term follow-up, the 5-year event-free survival (EFS) and 5-year overall survival (OS) of ALL patients were (63.1±3.3)% and (68.4±3.2)%. The 7-year EFS and OS were (61.0±3.5)% and (67.6±3.3)%.The 5-year EFS of intermediate risk group in ALL-2005 and ALL-2009 protocol were (73.6±6.1)% and (71.7±4.3)% with no difference (χ(2)=0.064, P=0.801). The 5-year EFS of high risk group in two protocols were (27.6±9.6)% and (33.9±9.3)%, showing no significant difference (χ(2)=0.296, P=0.586). Five years relapsed rate of two protocols were (33.8±5.7)% and (32.6±4.1)% with no difference (χ(2)=0.055, P=0.815). The mortalities were 36.8% and 29.8% separately (χ(2)=2.869, P=0.090). Univariate analysis indicated that age, male, risk, BCR/ABL translocation/t(9;22) and resistant to induction were risk prognostic factors in long-term survival (χ(2)=4.764, 4.796, 46.410, 9.560, 25.450; P=0.029, 0.029, <0.001, 0.049, <0.001). Cox multivariate analysis showed male, risk and resistant to induction were independent risk prognostic factors (RR=1.790, 2.727, 2.719; P=0.021, 0.000, 0.012). Conclusion: Protocol ALL-2009 enhanced the chemotherapy intensity in intermediate risk group with no benefit of survival. BCR-ABL fusion or t(9;22) translocation was still the risk factor of prognosis. TKI inhibitor used in these patients could improve survival. EFS rate was increased a little and death rate was decreased in ALL-2009 protocol with no significant lower relapsed rate comparing with ALL-2005 protocol.

[Long-term results of multicenter study based on childhood acute lymphoblastic leukemia 2005 protocol].

Objective: To evaluate the long-term efficacy and prognostic factors of childhood acute lymphoblastic leukemia (ALL) enrolled in Shanghai Children's Medical Center-Acute Lymphoblastic Leukemia-2005(SCMC-ALL-2005) multicenter study. Methods: Between May 2005 and December 2014, 1 497 newly diagnosed ALL patients were enrolled and treated in 5 hospitals of SCMC-ALL-2005 study group, using risk-stratified SCMC-ALL-2005 protocol. Risk group classification and treatment intensity were based on clinical features, genetic abnormalities, early response to treatment and levels of minimal residual disease (MRD). Kaplan-Meier method was used to generate overall survival (OS) and event-free survival(EFS) curves. Cox proportional hazards models were used for multivariate analyses. Results: The patients were followed up to December 31, 2016, the median follow-up time was 69 months (24-141 months). The 5-year and 10-year OS rates were (80.0±1.0)% and (76.0±2.0)%. The 5-year and 10-year EFS rates were (69.0±1.0)% and (66.0±2.0)%. The 5-year and 10-year relapse rates were (23.0±1.0)% and (25.0±2.0)%. The 5-year OS and EFS for low risk (LR), intermediate risk (IR) and high risk (HR) were (91.1±1.4)% and (83.3±1.8)%, (79.2±1.5)% and (68.9±1.7)%, (52.9±4.4)% and (30.0±3.8)%, respectively. MRD negative status (<0.01%) on day 55 was seen in 792 patients (82.8%) and positive MRD on day 55 was associated with poor prognosis (OR=1.9, 95%CI: 1.3-2.7, P=0.001). Twenty-four HR patients received allogeneic hematopoietic stem cell transplantation and 17(70.8%) of them were alive and in remission. A total of 164 severe adverse events occurred, 46 of them died, treatment-related mortality was 3.1%. Conclusions: In this large sample research, the overall outcome for multi-center SCMC-ALL-2005 study was favorable. This helps to promote the standardized treatment of childhood ALL to the whole country. MRD results on day 55 of induction therapy have important prognostic and therapeutic implications.

Molecular response in acute promyelocytic leukemia: a direct comparison of regular and real-time RT-PCR.

Evaluation of molecular response is important for the diagnosis and monitoring of minimal residual disease in patients with acute promyelocytic leukemia (APL). In this study, we analyzed the molecular response by regular reverse transcription-polymerase chain reaction (RT-PCR) and quantitative real-time RT-PCR in 31 newly diagnosed patients. The real-time RT-PCR results are reported as normalized DoseN and log-reduction (3.0-4.9 log-reduction as minor and > or =5.0 log-reduction as major molecular response). After induction therapy and completion of consolidation, minor molecular response was documented in 35.5 and 96.8% patients, respectively, which was equivalent to the regular RT-PCR (22.6 and 96.8%), whereas the major molecular response rate was significantly lower (12.9 and 90.3%, respectively). All patients achieved major molecular response during and after maintenance therapy. During the follow-up study, loss of major molecular response was observed in two patients, which was associated with subsequent loss of minor molecular response, positive RT-PCR and then documentation of central nervous system leukemia or clinical relapse in 3-6 months. For summary, we demonstrated that the real-time RT-PCR is potentially superior to regular RT-PCR in evaluation of molecular response in APL patients and that reporting real-time RT-PCR data by log-reduction is feasible and clinically relevant.

A role for the SHP-2 tyrosine phosphatase in nerve growth-induced PC12 cell differentiation.

SHP-1 and SHP-2 are intracellular protein tyrosine phosphatases containing two adjacent src homology 2 domains that target these phosphatases to cell surface receptor signaling complexes and play a role in receptor signal transduction. In this report the PC12 cell system was used to investigate the potential roles of SHP-1 and SHP-2 in the induction of neuronal differentiation by nerve growth factor (NGF). By using neurite outgrowth as a marker for differentiation, the effects of transfected constructs of SHP-1 and SHP-2 were assessed. Overexpression of a catalytically inactive SHP-2, but not a catalytically inactive SHP-1, blocked NGF-stimulated neurite outgrowth. The mitogen-activated protein kinase (MAPK) signaling cascade is important for the morphological differentiation in PC12 cells, and both SHP-1 and SHP-2 have been implicated to act upstream of MAPK in other receptor signaling systems. A positive role for SHP-2 but not SHP-1 in the activation of MAPK by NGF was demonstrated by introduction of the SHP-2 phosphatase mutants along with hemagglutinin-tagged MAPK. Coexpression studies with the SHP-2 mutant along with mutant forms of MAPK kinase suggested that SHP-2 functions upstream of MAPK kinase and MAPK in NGF-induced neurite outgrowth.

Interdependent regulation of intracellular acidification and SHP-1 in apoptosis.

The G protein-coupled receptor agonist somatostatin (SST)-induces apoptosis in MCF-7 human breast cancer cells. This is associated with induction of wild-type p53, Bax, and an acidic endonuclease. We have shown recently that its cytotoxic signaling is mediated via membrane-associated SHP-1 and is dependent on decrease in intracellular pH (pHi) to 6.5. Here we investigated the relationship between intracellular acidification and SHP-1 in cytotoxic signaling. Clamping of pHi at 7.25 by the proton-ionophore nigericin abolished SST-signaled apoptosis without affecting its ability to regulate SHP-1, p53, and Bax. Apoptosis could be induced by nigericin clamping of pHi to 6.5. Such acidification-induced apoptosis was not observed at pHi <6.0 or >6.7. pHi-dependent apoptosis was associated with the translocation of SHP-1 to the membrane, enhanced in cells overexpressing SHP-1, and was abolished by its inactive mutant SHP-1C455S. Acidification caused by inhibition of Na+/H+ exchanger and H+ ATPase (pHi = 6.55 and 6.65, respectively) also triggered apoptosis. The effect of concurrent inhibition of Na+/H+ exchanger and H(+)-ATPase on pHi and apoptosis was comparable with that of SST. Acidification-induced, SHP-1-dependent apoptosis occurred in breast cancer cell lines in which SST was cytotoxic (MCF-7 and T47D) or not (MDA-MB-231). We conclude that: (a) SST-induced SHP-1-dependent acidification occurs subsequent to or independent of the induction of p53 and Bax; (b) SST-induced intracellular acidification may arise due to inhibition of Na+/H+ exchanger and H(+)-ATPase; and (c) SHP-1 is necessary not only for agonist-induced acidification but also for the execution of acidification-dependent apoptosis. We suggest that combined targeting of SHP-1 and intracellular acidification may lead to a novel strategy of anticancer therapy bypassing the need for receptor-mediated signaling.

[Outcome of childhood T-cell acute lymphoblastic leukemia: a report of 99 cases].

OBJECTIVE: To explore the incidence, clinical characteristics and prognosis of childhood T-cell acute lymphoblastic leukemia (T-ALL) enrolled in protocol ALL-2005. METHOD: Retrospective reviews of patients diagnosed as having T-ALL were studied in the period between May 2005 and August 2013 in Shanghai Children's Medical Center. Predictive values of early treatment responses, including prednisone response, bone marrow morphology on day 35 during induction chemotherapy, and minimal residual disease (MRD) monitored by flow cytometry on day 35 and 55, were analyzed. The Kaplan-Meier method was used to assess the survival rates. RESULT: Ninety-nine evaluable patients aged 1-18 years with newly diagnosed T-ALL were enrolled in protocol ALL-2005. The median follow-up period for patients remaining alive in continuous remission was 60 months (24-109 months). The 5-year event free survival (EFS) and overall survival (OS) rates for these patients were 65% and 69%, respectively. Forty-three (43%) patients were older than 10 years at diagnosis. A total of 61 patients presented with initial white blood cell count (WBC)≥50×10(9)/L. Thirty-seven patients with mediastinal mass were found in this study. Seventy-one patients were defined as prednisone good responders (PGR) on day 8 of prednisone therapy. Ninty-four patients achieved complete remission (CR) on day 35 of induction therapy. MRD negative status (<0.01%) on day 35 were seen in 55% of the 42 assessed patients. MRD negative status on day 55 was seen in 78% of the 52 assessed patients. Patients with high MRD (>1%) on day 55 5-year EFS was 40% were associated with poor prognosis (P=0.03). Early T-cell precursor (ETP) subtype was not associated with treatment outcome in this study. Six patients abandoned therapy, 10 lost to follow-up, 22 had relapsed disease, 1 was diagnosed as having a second tumor and 20 patients died. CONCLUSION: Overall outcome for T-ALL on protocol ALL-2005 is favorable. MRD results on day 55 of induction therapy have important prognostic and therapeutic implications.

Octapeptide somatostatin analog SMS 201-995 induces translocation of intracellular PTP1C to membranes in MCF-7 human breast adenocarcinoma cells.

Somatostatin (SST) analogs exert direct antiproliferative actions in pancreatic, pituitary, and mammary tumor cells in vitro. SST receptor (SSTR)-mediated induction of membrane-associated protein tyrosine phosphatase (PTP) activity has been implicated in its anti-proliferative signaling by virtue of its ability to dephosphorylate and inactivate growth factor receptor kinases. Recently, a PTP-containing Src homology 2 domain, identified as PTP1C/SHPTP1/SHP/HCP, was found to be associated with SSTR in rat pancreatic acinar cell membranes. In the present study we investigated the antiproliferative action of the octapeptide SST analog SMS 201-995 (OCT) and its effect on PTP activity in MCF-7 human breast adenocarcinoma cells. We report here that OCT does not directly stimulate membrane-associated PTP activity, but induces translocation of intracellular PTP to the membrane in MCF-7 cells preincubated with the peptide in a time- and concentration-dependent manner. We demonstrate that this is due at least in part to OCT-induced recruitment of cytosolic PTP1C. OCT-induced recruitment of PTP1C to the cell surface as well as its ability to inhibit the growth of MCF-7 cells was G protein dependent and inhibited by orthovanadate. These findings suggest that translocation of cytosolic PTP1C by SST analogs to the cell surface is an early event in its antiproliferative signaling in tumor cells.