MAFF mediates PEITC-induced enhancement of sensitivity to carboplatin in ovarian cancer cell lines via activating ZNF711 transcription in vivo and invitro.

Enhanced drug resistance poses a significant challenge in treating ovarian cancer (OC). Phenylethyl isothiocyanate (PEITC) is involved in drug resistance in OC, but the mechanism remains unclear. In this study, we investigated the molecular regulatory mechanism of carboplatin sensitivity in OC associated with PEITC, MAF BZIP Transcription Factor F (MAFF), and Zinc finger proteins (ZNF) 711. The carboplatin sensitivity was significantly increased in OC cells after PEITC treatment. Knockdown of MAFF significantly enhanced the carboplatin sensitivity of OC cells, promoted apoptosis, inhibited colony-forming efficiency in vitro, and suppressed tumor growth in vivo. The binding site of MAFF to the ZNF711 promoter was predicted, and the knockdown of MAFF significantly increased the ZNF711 expression. Results of the dual luciferase assay and ChIP-PCR confirmed the binding of MAFF to the ZNF711 promoter. Immunofluorescence and CoIP results demonstrated the colocalization and the binding of MAFF and its interacting protein, BZIP Transcription Factor ATF-like 3 (BATF3). Similarly, we confirmed the binding of BATF3 to the ZNF711 promoter. Knockdown of BATF3 alone and simultaneous knockdown of BATF3 and MAFF showed similar regulatory effects on ZNF711 transcription and apoptosis. These suggested that the binding of MAFF to BATF3 inhibited ZNF711 transcription and reduced carboplatin sensitivity in OC.

Distributed Discrete-Time Convex Optimization With Closed Convex Set Constraints: Linearly Convergent Algorithm Design.

The convergence rate and applicability to directed graphs with interaction topologies are two important features for practical applications of distributed optimization algorithms. In this article, a new kind of fast distributed discrete-time algorithms is developed for solving convex optimization problems with closed convex set constraints over directed interaction networks. Under the gradient tracking framework, two distributed algorithms are, respectively, designed over balanced and unbalanced graphs, where momentum terms and two time-scales are involved. Furthermore, it is demonstrated that the designed distributed algorithms attain linear speedup convergence rates provided that the momentum coefficients and the step size are appropriately selected. Finally, numerical simulations verify the effectiveness and the global accelerated effect of the designed algorithms.

Cruciferous vegetable consumption and multiple health outcomes: an umbrella review of 41 systematic reviews and meta-analyses of 303 observational studies.

Background: Epidemiological studies evaluating the associations between the consumption of cruciferous vegetables (CV) and diverse health outcomes have generated inconsistent findings. Therefore, we carried out an umbrella review to systematically summarize existing evidence on this topic. Methods: This study had been registered at PROSPERO (no. CRD42021262011). Relevant systematic reviews and meta-analyses of observational studies were identified by searching PubMed, Web of science, and Embase databases from inception up to March 15, 2021. Observational studies investigating the association between CV intake and multiple health outcomes in humans were eligible for inclusion. The validated AMSTAR (A Measurement Tool to Assess Systematic Reviews) instrument was utilized for assessing the methodological quality of the included systematic reviews. For each meta-analysis, we assessed the summary effect size by using fixed and random effects models, 95% prediction intervals, heterogeneity, evidence of small-study effects, and excess significance bias. Results: Our umbrella review included 41 meta-analyses of 303 individual studies involving 13 394 722 participants. Twenty-four health outcomes including cancers (n = 23), cardiovascular disease (n = 12), mortality (n = 5), and metabolic diseases (n = 1) were evaluated. The summary random effects estimates were significant at P < 0.05 in 24 meta-analyses - all of which reported decreased risks of health outcomes. All were of moderate methodological quality in our study. Of the 41 meta-analyses, we observed suggestive evidence for beneficial associations between gastric cancer, lung cancer, endometrial cancer, and all-cause mortality. Moreover, 16 associations were supported by weak evidence, including breast cancer, lung cancer, renal cell carcinoma, bladder cancer, prostate cancer, ovarian cancer, endometrial cancer, colon cancer, colorectal adenoma, colorectal neoplasm, non-Hodgkin lymphoma, and total cancer. Conclusions: It revealed that CV intake might be associated with beneficial effects on several health-related outcomes (gastric cancer, lung cancer, endometrial cancer, and all-cause mortality). Other associations could be genuine, but substantial uncertainty remains. Additional studies are needed to evaluate the relationship between the consumption of CV and various health outcomes as well as robust randomized controlled trials in the future.

Pre-diagnosis Cruciferous Vegetables and Isothiocyanates Intake and Ovarian Cancer Survival: A Prospective Cohort Study.

Background: The associations of the consumption of cruciferous vegetables (CVs) and their bioactive components, isothiocyanates (ITCs), with ovarian cancer (OC) mortality have been unclear, owing to limited studies and inconsistent findings. To date, no studies have evaluated these associations among Chinese patients with OC. This study aims to provide more evidence indicating the relationships of pre-diagnosis CVs and ITC intake with OC survival. Methods: We examined the associations of pre-diagnosis CV and ITC intake with OC mortality in a hospital-based cohort (n = 853) of Chinese patients with epithelial OC between 2015 and 2020. Pre-diagnosis dietary information was evaluated with a validated food frequency questionnaire. Deaths were ascertained until March 31, 2021 via medical records and active follow-up. The associations were examined with the Cox proportional hazards model, adjusted for potential confounders, and stratified by menopausal status, residual lesions, histological type, and body mass index (BMI). Results: During a median follow-up of 37.2 months (interquartile: 24.7-50.2 months), we observed 130 deaths. The highest tertile of total CV intake was associated with better survival than the lowest tertile intake [hazard ratio (HR) = 0.57, 95% confidence interval (CI) = 0.33-0.98, p trend < 0.05]. In addition, higher intake of ITCs from CVs was associated with better survival (HRT3VS.T1 = 0.59, 95% CI = 0.36-0.99, p trend = 0.06). Significant inverse associations were also observed for subgroup analyses stratified by menopausal status, residual lesions, histological type, and BMI, although not all associations showed statistical significance. Conclusion: Increasing pre-diagnosis consumption of CVs and ITCs was strongly associated with better survival in patients with OC.

miR-19-3p Promotes Autophagy and Apoptosis in Pelvic Organ Prolapse Through the AKT/mTOR/p70S6K Pathway: Function of miR-19-3p on Vaginal Fibroblasts by Targeting IGF-1.

OBJECTIVE: Pelvic organ prolapse (POP) is a common condition in older women. A decrease in collagen 1 (Col-1) expression is one of the main causes of POP. Many microRNAs play an important role in regulating target genes. The relationship between miR-19-3p and POP is investigated in this study, and the molecular mechanism was also explored to find whether miR-19-3p may be a potential target for early diagnosis and prevention of POP. METHODS: A total of 60 patients with POP and 60 patients without POP were included in this study. Reverse transcription-polymerase chain reaction and Western blot were used to detect the expression of miR-19-3p, insulin-like growth factor 1 (IGF-1), and the Akt/mTOR/p70S6K pathway. Cell cycle was defined by flow cytometric analysis. The combination of miR-19-3p and IGF-1 was revealed by luciferase assays. RESULTS: The results of this study show that miR-19-3p was upregulated in the tissue of patients with POP, whereas COL-1 and IGF-1 expressions were lower in the POP group. miR-19-3p promoted excessive fibroblast autophagy and apoptosis. miR-19-3p negatively regulated the Akt/mTOR/p70S6K pathway and inhibited COL-1 secretion. Luciferase reporter assay showed that miR-19-3p regulated IGF-1 expression by direct target binding. CONCLUSIONS: miR-19-3p has negative associations with the expression of Col-1. Our study highlights that miR-19-3p may affect the synthesis of Col-1 by targeting IGF-1 and that it may play an vital role in POP.

The role of MMP-2 and MMP-9 in the metastasis and development of hypopharyngeal carcinoma.

INTRODUCTION: The role of matrix metalloproteinase-2 and 9 in the metastasis and development of hypopharyngeal carcinoma has not been clarified. OBJECTIVES: To observe the relationship between matrix metalloproteinase-2, matrix metalloproteinase-9 and the metastasis, development of hypopharyngeal carcinoma. METHODS: This study included 42 hypopharyngeal cancer patients. The mRNA and protein expression levels of matrix metalloproteinase-2 and 9 in hypopharyngeal carcinoma and paracancerous tissues were detected by reverse transcription-polymerase chain reaction and Western blot. RESULTS: Reverse transcription-polymerase chain reaction detection showed that the mRNA of matrix metalloproteinase-2 and 9 was expressed in both cancer and pericarcinoma tissues, but was almost not expressed in polypoid control tissues. The expression intensity in the cancer tissue was significantly higher than that in the pericarcinoma tissue (matrix metalloproteinase-2: t ＝ 2.529, p = 0.015; matrix metalloproteinase-9: t ＝ 4.781, p ＜ 0.001). The mRNA expression in the cancer tissue was enhanced with the increase of the tumor clinical stage (matrix metalloproteinase-2: F = 4.003, p = 0.026; matrix metalloproteinase-9: F = 5.501, p = 0.008). Its expression intensity was associated with the metastasis of lymph nodes (N staging) and increased with the degree of lymphatic metastasis (matrix metalloproteinases-2: F = 8.965, p = 0.005; matrix metalloproteinase-9: F = 5.420, p = 0.025). There was no significant change in T staging of tumor. With the increase of tumor pathological stage, the mRNA expression of matrix metalloproteinase-2 and 9 was strengthened (matrix metalloproteinase-2: F = 3.884, p = 0.029; matrix metalloproteinase-9: F = 3.783, p = 0.032). The protein expression level of matrix metalloproteinase-2 and 9 was the same as that of mRNA. CONCLUSION: The expression of matrix metalloproteinase-2 and 9 in hypopharyngeal carcinoma was significantly higher than that in pericarcinoma tissue, and it was enhanced with the increase of clinical stage. The expression level was related to lymph node metastasis and tumor pathological stage. Thus, matrix metalloproteinase-2 and 9 may be involved in the occurrence, development, invasion and metastasis of hypopharyngeal carcinoma through a variety of mechanisms.

Improvement in Outlet Obstructive Constipation Symptoms After Vaginal Stent Treatment for Rectocele.

Objective. The objective is to determine the possible improvement in outlet obstructive constipation symptoms after vaginal stent treatment for rectocele. Methods. Female patients with rectocele (n = 156) accompanied with outlet obstructive constipation were selected in this study. Longo's obstructed defecation syndrome (ODS) questionnaire, rectoanal pressures, and rectal balloon expulsion (BET) were evaluated at baseline, 1 month follow-up, and 6 months follow-up. Moreover, the side effects and the potential reasons for giving up treatment were also detected. Results. Vaginal stent significantly decreased the straining intensity, shortened the straining extensity time, decreased the use of laxatives, and alleviated the symptoms of incomplete evacuation (P < .05). The vaginal stent also increased the rectal pressure and shortened the balloon expulsion time (P < .05). Conclusions. As an effective, feasible, and safe procedure, the vaginal stent can be recommended as a treatment of choice for rectocele combined with outlet obstructive constipation.

TIPRL, a Novel Tumor Suppressor, Suppresses Cell Migration, and Invasion Through Regulating AMPK/mTOR Signaling Pathway in Gastric Cancer.

Invasion and metastasis of gastric cancer after curative resection remain the most common lethal outcomes. However, our current understanding of the molecular mechanism underlying gastric cancer metastasis is far from complete. Herein, we identified TOR signaling pathway regulator (TIPRL) as a novel metastasis suppressor in gastric cancer through genome-wide gene expression profiling analysis using mRNA microarray. Decreased TIPRL expression was detected in clinical gastric cancer specimens, and low TIPRL expression was correlated with more-advanced TNM stage, distant metastasis, and poor clinical outcome. Moreover, TIPRL was identified as a direct target of miR-216a-5p and miR-383-5p. Functional study revealed that re-expression of TIPRL in gastric cancer cell lines suppressed their migratory and invasive capacities, whereas inverse effects were observed in TIPRL-deficient models. Mechanistically, TIPRL downstream effectors and signaling pathways were investigated using mRNA microarray. Gene expression profiling revealed that TIPRL could not modulate the downstream genes at transcriptional levels, thereby implying that the regulation might occur at the post-transcriptional levels. We further demonstrated that TIPRL induced phosphorylation/activation of AMPK, which in turn attenuated phosphorylation of mTOR, p70S6K, and 4E-BP1, thereby leading to inactivation of mTOR signaling and subsequent suppression of cell migration/invasion in gastric cancer. Taken together, TIPRL acts as a novel metastasis suppressor in gastric cancer, at least in part, through regulating AMPK/mTOR signaling, likely representing a promising target for new therapies in gastric cancer.

Risk Factors for Postoperative Recurrence of Chronic Suppurative Otitis Media / 中国医学科学院学报

To analyze the risk factors for postoperative recurrence of chronic suppurative otitis media(CSOM) and explore the intervention measures to prevent postoperative recurrence of CSOM. A total of 1066 patients with CSOM who underwent concurrent surgical treatment and achieved clinical cure in our hospital from January 2012 to December 2018 were enrolled.The clinical data and laboratory findings were reviewed by using an electronic medical record system and the patients were followed up for 1 year.The patients were divided into the non-recurrent group and the recurrent group.Chi-square test and multivariate logistic regression were used to compare the factors may contribute to the postoperative recurrence. The recurrence rate of CSOM was 6.38%.Multi-drug-resistant(MDR) infection before surgery(=16.338,=0.000),aged ≥60 years(=5.182,=0.023),frequency of occurrence ≥3 times/year(=4.388,=0.036),duration of active period>7 d(=4.729,=0.030),repeated upper respiratory tract infection>3 times/year(=11.913,=0.001),accompanied by chronic sinusitis(=11.077,=0.001),blood glucose>6.11 mmol/L(=15.327,=0.000),postoperative serum procalcitonin(PCT)>0.5 μg/L(=8.337,=0.004) were the risk factors for postoperative recurrence.The use of snorkel was a protective factor for postoperative recurrence(=5.308,=0.021).Multivariate analysis showed that MDR infection(=3.373,95%:1.825-6.234,=0.000),repeated upper respiratory tract infection>3 times/year(=2.727,95%:1.479-5.030,=0.001),accompanied by chronic sinusitis(=2.980,95%:1.654-5.369,=0.000),blood glucose>6.11 mmol/L(=3.219,95%:1.741-5.953,=0.000),and postoperative serum PCT>0.5 μg/L(=2.085,95%:1.106-3.931,=0.023) were independent risk factors for postoperative recurrence in CSOM patients. Effective prevention and control of MDR infection,control of blood sugar,prevention of upper respiratory tract infection,and lowering the recurrence of chronic sinusitis are the main measures to reduce postoperative recurrence of CSOM.Monitoring of the infection marker PCT can help to achieve early intervention.

Prognostic Factors of Central Venous Catheter-related Bloodstream Infections / 中国医学科学院学报

Objective To explore the prognostic factors of central venous catheter-related bloodstream infection(CR-BSI)and provide reference for clinical practice. Methods The clinical data of 346 CR-BSI patients from February 2014 to July 2019 were retrospectively reviewed,and the prognostic factors were analyzed. Results Of the 346 CR-BSI patients,62 died,yielding a case-fatality rate of 17.92%.Univariate analysis showed that 18 factors including age(

Three-Dimensional Transperineal Ultrasonography for Diagnosis of Female Occult Stress Urinary Incontinence.

BACKGROUND We evaluated the utility of three-dimensional transperineal ultrasonography in detecting occult stress urinary incontinence in women undergoing anterior pelvic floor reconstruction surgery for severe cystocele. MATERIAL AND METHODS We enrolled 207 women with stage III-IV cystocele without urinary stress incontinence. One week before the operation, the patients underwent pelvic floor ultrasonography. We measured the vertical distance between the bladder neck and posterior margin of the pubic symphysis, the posterior vesicourethral angle, the urethral rotation angle, the formation of funnel shape, the hiatus area, and the length of the urethra and the funnel shape. Postoperatively, the patients were evaluated for symptoms of stress urinary incontinence and with the 20-minute pad test. RESULTS The posterior vesicourethral angle with Valsalva maneuver, the difference in the posterior vesicourethral angle between the resting state and with the Valsalva state, and the angle of the proximal urethra were larger in the incontinence-positive group than in the incontinence-negative group (P<0.05). Funnel shape urethra was longer in the incontinence-positive group than in the incontinence-negative group (P<0.05). The cutoff value was 137.5° for the posterior vesicourethral angle with Valsalva maneuver, 39.5° for the difference in the posterior vesicourethral angle, 44.5° for the angle of the proximal urethra, and 0.35 cm for the length of the funnel shape. Multivariate analysis revealed that the difference between the posterior vesicourethral angle in the resting state and with Valsalva, the angle of the proximal urethra, and the length of funnel shape were strongly correlated with occult stress urinary incontinence. CONCLUSIONS Ultrasonography is an effective method for identifying occult stress urinary incontinence.

Correlation between Asymptomatic Bacteriuria and Surgical Site Infection in Middle-aged and Elderly Patients Undergoing Open Hysterectomy / 中国医学科学院学报

Objective To explore the correlation between asymptomatic bacteriuria(AB)and surgical site infection(SSI)in middle-aged and elderly women undergoing open hysterectomy.Methods The clinical data of 1469 middle-aged and elderly women undergoing open hysterectomy in the Third Affiliated Hospital of Guizhou Medical University from June 2011 to August 2018 were retrospectively analyzed.Factors associated with SSI after operation were analyzed by univariate and multivariate regression models to identify the relationship of AB with SSI after open hysterectomy.Results Of these 1469 patients,101(6.88%)had SSI and 124 had AB[including 14 patients(11.29%)with infections].In addition,1345 patients had no AB,among whom 87(6.47%)had infections.Thus,the infection rate in patients with AB was significantly higher than that in patients without AB(=4.123,=0.042).Univariate analysis showed AB,history of diabetes mellitus,surgical procedure,length of stay(>15 d),season(summer and autumn),body mass index(≥25 kg/m ),nature of lesions(malignant tumors),ASA grade(>grade Ⅱ),incision length(≥10 cm),and operative time(≥3 h),bleeding volume(≥1000 ml),serum albumin concentration(grade Ⅱ)were risk factors for SSI in these patients(all <0.05). Conclusions AB is one of the risk factors for SSI in middle-aged and elderly women undergoing open hysterectomy.Screening and treatment of AB before surgery can reduce the risk of SSI.ASA grading shall be performed before surgery before corresponding preparation was offered.Effective control of blood glucose,improved surgical skills,and shorter operative time are helpful for lowering postoperative SSI.

The Expression of SOCS and NF-Ï°B p65 in Hypopharyngeal Carcinoma.

BACKGROUND: Hypopharyngeal carcinoma is one of the most common types of head and neck tumors. Suppressers of cytokine signalling (SOCS) family members are key regulators of cytokine homeostasis, they play important roles in the process of cell proliferation, differentiation, maturation and apoptosis, and participate in the occurrence and development of tumor. The abnormal activation of NF-Ï°B is an important feature of the tumor. The aim of this study was to investigate the relationships among SOCS, NF-Ï°B p65 and hypopharyngeal carcinoma development.C. METHODS: We included 72 hypopharyngeal cancer patients and 9 swallow cyst patients. The patients were recruited at The Second Hospital of Shandong University (Jinan, China) between 2014 and 2016. The mRNA and protein expression levels of SOCS-1, SOCS-3 and NF-Ï°B p65 in hypopharyngeal carcinoma tissues, para-cancerous tissues and control tissues were detected by RT-PCR and Western blot analysis, respectively. RESULTS: Hypopharyngeal carcinoma tissues had lower level expression of SOCS-1 and SOCS-3 than pericarcinoma tissues, but there was no significant difference, while cancer tissues had significantly higher level expression of NF-Ï°B p65 than that of pericarcinoma tissues (0.412±0.266, 0.281±0.231, t=2.969, P=0.004). The early stage patients had striking higher level expression of SOCS-1 and SOCS-3 than that in advanced stages (F=16.202, P<0.001; F=52.295, P<0.001), while the expression of NF-Ï°B p65 in early stages had lower level than that in advanced stages (F=3.383, P=0.04). CONCLUSION: SOCS-1, SOCS-3 may be protective factors while NF-Ï°B p65 could be a harmful factor in hypopharyngeal carcinoma.

Correction: PD-1 Blockade and OX40 Triggering Synergistically Protects against Tumor Growth in a Murine Model of Ovarian Cancer.

[This corrects the article DOI: 10.1371/journal.pone.0089350.].

A Single Nucleotide Variant in HNF-1ß is Associated with Maturity-Onset Diabetes of the Young in a Large Chinese Family.

BACKGROUND: Maturity-onset diabetes of the young (MODY) is a heterogeneous entity of monogenic disorders characterized by autosomal dominant inheritance. Eleven genes were related, including HNF4α, GCK, HNF1α, IPF1, and HNF-1ß, and various mutations are being reported. METHODS: To help the overall understanding of MODY-related pathologic mutations, we studied a large MODY family found in 2012, in Shandong, China, which contained 9 patients over 3 generations. DNA was extracted from the periphery blood samples of (i) 9 affected members, (ii) 17 unaffected members, and (iii) 1000 healthy controls. Three pooled samples were obtained by mixing equal quantity of DNA of each individual within the each group. Totally 400 microsatellite markers across the whole genome were genotyped by capillary electrophoresis. The known MODY-related gene near the identified marker was sequenced to look for putative risk variants. RESULTS: Allelic frequency of marker D17S798 on chromosome 17q11.2 were significantly different (P<0.001) between the affected vs. unaffected members and the affected vs. healthy controls, but not between the unaffected members vs. healthy controls. MODY5-related gene, hepatocyte nuclear factor-1ß (HNF-1ß) on 17q12 near D17S798 became the candidate gene. A single nucleotide variant (SNV) of C77T in the non-coding area of exon 1 of HNF-1ß was found to be related to MODY5. CONCLUSION: This novel SNV of HNF-1ß contributes to the diabetes development in the family through regulating gene expression most likely. The findings help presymptomatic diagnosis, and imply that mutations in the non-coding areas, as well as in the exons, play roles in the etiology of MODY.

PD-1 blockade and OX40 triggering synergistically protects against tumor growth in a murine model of ovarian cancer.

The co-inhibitory receptor Programmed Death-1 (PD-1) curtails immune responses and prevent autoimmunity, however, tumors exploit this pathway to escape from immune destruction. The co-stimulatory receptor OX40 is upregulated on T cells following activation and increases their clonal expansion, survival and cytokine production when engaged. Although antagonistic anti-PD-1 or agonistic anti-OX40 antibodies can promote the rejection of several murine tumors, some poorly immunogenic tumors were refractory to this treatment. In the present study, we evaluated the antitumor effects and mechanisms of combinatorial PD-1 blockade and OX40 triggering in a murine ID8 ovarian cancer model. Although individual anti-PD-1 or OX40 mAb treatment was ineffective in tumor protection against 10-day established ID8 tumor, combined anti-PD-1/OX40 mAb treatment markedly inhibited tumor outgrowth with 60% of mice tumor free 90 days after tumor inoculation. Tumor protection was associated with a systemic immune response with memory and antigen specificity and required CD4(+) cells and CD8(+) T cells. The anti-PD-1/OX40 mAb treatment increased CD4(+) and CD8(+) cells and decreased immunosuppressive CD4(+)FoxP3(+) regulatory T (Treg) cells and CD11b(+)Gr-1(+) myeloid suppressor cells (MDSC), giving rise to significantly higher ratios of both effector CD4(+) and CD8(+) cells to Treg and MDSC in peritoneal cavity; Quantitative RT-PCR data further demonstrated the induction of a local immunostimulatory milieu by anti-PD-1/OX40 mAb treatment. The splenic CD8(+) T cells from combined mAb treated mice produced high levels of IFN-γ upon tumor antigen stimulation and exhibited antigen-specific cytolytic activity. To our knowledge, this is the first study testing the antitumor effects of combined anti-PD-1/OX40 mAb in a murine ovarian cancer model, and our results provide a rationale for clinical trials evaluating ovarian cancer immunotherapy using this combination of mAb.

Suppression effect of recombinant adenovirus vector containing hIL-24 on Hep-2 laryngeal carcinoma cells.

The melanoma differentiation-associated gene-7 [MDA-7; renamed interleukin (IL)-24] was isolated from human melanoma cells induced to terminally differentiate by treatment with interferon and mezerein. MDA-7/IL-24 functions as a multimodality anticancer agent, possessing proapoptotic, antiangiogenic and immunostimulatory properties. All these attributes make MDA-7/IL-24 an ideal candidate for cancer gene therapy. In the present study, the human MDA-7/IL-24 gene was transfected into the human laryngeal cancer Hep-2 cell line and human umbilical vein endothelial cells (HUVECs) with a replication-incompetent adenovirus vector. Reverse transcription polymerase chain reaction and western blot analysis confirmed that the Ad-hIL-24 was expressed in the two cells. The expression of the antiapoptotic gene, Bcl-2, was significantly decreased and the IL-24 receptor was markedly expressed in Hep-2 cells following infection with Ad-hIL-24, but not in HUVECs. In addition, the expression of the proapoptotic gene, Bax, was induced and the expression of caspase-3 was increased in the Hep-2 cells and HUVECs. Methyl thiazolyl tetrazolium assay indicated that Ad-hIL-24 may induce growth suppression in Hep-2 cells but not in HUVECs. In conclusion, Ad-hIL-24 selectively inhibits proliferation and induces apoptosis in Hep-2 cells. No visible damage was found in HUVECs. Therefore, the results of the current study indicated that Ad-hIL-24 may have a potent suppressive effect on human laryngeal carcinoma cell lines, but is safe for healthy cells.

Combined TIM-3 blockade and CD137 activation affords the long-term protection in a murine model of ovarian cancer.

BACKGROUND: T-cell immunoglobulin and mucin domain 3 (TIM-3) is known as a negative immune regulator and emerging data have implicated TIM-3 a pivotal role in suppressing antitumor immunity. The co-stimulatory receptor CD137 is transiently upregulated on T-cells following activation and increases their proliferation and survival when engaged. Although antagonistic anti-TIM-3 or agonistic anti-CD137 antibodies can promote the rejection of several murine tumors, some poorly immunogenic tumors were refractory to this treatment. In this study, we sought to evaluate whether combined TIM-3 blockade and CD137 activation would significantly improve the immunotherapy in the murine ID8 ovarian cancer model. METHODS: Mice with established ID8 tumor were intraperitoneally injected with single or combined anti-TIM-3/CD137 monoclonal antibody (mAb); mice survival was recorded, the composition and gene expression of tumor-infiltrating immune cells in these mice was analyzed by flow cytometry and quantitative RT-PCR respectively, and the function of CD8⁺ cells was evaluated by ELISA and cytotoxicity assay. RESULTS: Either anti-TIM-3 or CD137 mAb alone, although effective in 3 days established tumor, was unable to prevent tumor progression in mice bearing 10 days established tumor, however, combined anti-TIM-3/CD137 mAb significantly inhibited the growth of these tumors with 60% of mice tumor free 90 days after tumor inoculation. Therapeutic efficacy was associated with a systemic immune response with memory and antigen specificity, required CD4⁺ cells and CD8⁺ cells. The 2 mAb combination increased CD4⁺ and CD8⁺ cells and decreased immunosuppressive CD4⁺FoxP3⁺ regulatory T (Treg) cells and CD11b⁺Gr-1⁺ myeloid suppressor cells (MDSC) at tumor sites, giving rise to significantly elevated ratios of CD4⁺ and CD8⁺ cells to Treg and MDSC; This is consistent with biasing local immune response towards an immunostimulatory Th1 type and is further supported by quantitative RT-PCR data showing the increased Th1-associated genes by anti-TIM-3/CD137 treatment. The increased CD8⁺ T cells produced high level of IFN-γ upon tumor antigen stimulation and displayed antigen-specific cytotoxic activity. CONCLUSIONS: To our knowledge, this is the first report investigating the effects of anti-TIM-3/CD137 combined mAb in a murine ovarian cancer model, and our results may aid the design of future trials for ovarian cancer immunotherapy.

Lack of association between NCAM1 and early onset schizophrenia in a family based study in Shandong peninsula of China.

The neural cell adhesion molecule (NCAM1) gene plays important roles in cellular migration, synaptic integrity and neurodevelopment. Multiple NCAM1 proteins are differentially altered in schizophrenia (SZ). A whole genome association study was first carried out on Affymetrix genome-wide human single-nucleotide polymorphism (SNP) Array 6.0 and two pooled DNA samples consisting of 89 early onset SZ (EOS) cases and 1,000 controls. Association between rs10891495 and EOS was detected (χ(2) = 2 3.66, P = 1.15E-06). The position of this SNP is just within the NCAM1 gene. Since several previous studies reported that NCAM1 was a candidate gene for SZ, we further performed a family based association study and genotyped six SNPs (rs10891495, rs1245133, rs1821693, rs686050, rs12794326, rs674246) within NCAM1 gene in 100 EOS nuclear families. We found no evidence for association with SZ status either for SNP or for haplotype. Therefore, the NCAM1 gene is unlikely to play a major role in the etiology of early-onset SZ in the Chinese population.

Whole genome association study in a homogenous population in Shandong peninsula of China reveals JARID2 as a susceptibility gene for schizophrenia.

DNA pooling can provide an economic and efficient way to detect susceptibility loci to complex diseases. We carried out a genome screen with 400 microsatellite markers spaced at approximately 10 cm in two DNA pools consisting of 119 schizophrenia (SZ) patients and 119 controls recruited from a homogenous population in the Chang Le area of the Shandong peninsula of China. Association of D6S289, a dinucleotide repeat polymorphism in the JARID2 gene with SZ, was found and confirmed by individual genotyping (X(2) = 17.89; P = .047). In order to refine the signal, we genotyped 14 single nucleotide polymorphisms (SNPs) covering JARID2 and the neighboring gene, DNTBP1, in an extended sample of 309 cases and 309 controls from Shandong peninsula (including the samples from the pools). However, rs2235258 and rs9654600 in JARID2 showed association in allelic, genotypic and haplotypic tests with SZ patients from Chang Le area. This was not replicates in the extended sample, we conclude that JARID2 could be a susceptibility gene for SZ.