[Effects of Toxoplasma gondii infection on mouse and human uterine natural killer cells].

OBJECTIVE: To examine the effects of Toxoplasma gondii infection on the proportion, quantity, differentiation and function of mouse and human uterine natural killer cells (uNK cells), so as to explore the role of uNK cells in abortion of early pregnancy caused by T. gondii infection. METHODS: Pregnant mice were injected intraperitoneally with T. gondii tachyzoites on day 6.5 of pregnancy, and the abortion mouse model caused by T. gondii infections was constructed. Mouse uterine lymphocytes were isolated on day 9.5 of pregnancy. Human uterine lymphocytes were isolated from fresh human decidual specimens after abortion in normal early pregnancy and co-cultured with tachyzoites of the T. gondii RH strain for 48 h at T. gondii/uterine lymphocytes ratios of 0.5:1, 1:1 and 2:1. The phenotypes of mouse uNK cells (CD122, NK1.1, DX5) and human uNK cells (CD3, CD56, CD11b, CD27) and the expression of intracellular cytokines interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were detected by flow cytometry. Mouse and human uNK cells were sorted by magnetic beads, and the cytotoxicity of uNK cells was tested using the lactate dehydrogenase (LDH) release assay at effector/target cell ratios of 1:1, 5:1, 10:1 and 20:1 with mouse or human uNK cells as effector cells and mouse YAC-1 cells or human K562 cells as target cells. RESULTS: On day 9.5 of pregnancy, the mouse abortion rate was significantly higher in the infected group than that in the control group (83.02% vs. 3.51%; χ2 = 71.359, P < 0.001). Significantly lower absolute number of uNK cells [(4 547 ± 1 610) cells/mouse vs. (8 978 ± 3 339) cells/mouse; U = 2.000, P < 0.05], lower NK1.1 expression on uNK cell surface [(74.53 ± 8.37)% vs. (93.00 ± 1.11)%; U = 0.000, P < 0.05], higher proportion of NK1.1-DX5-cells [(20.10 ± 8.03)% vs. (5.04 ± 0.68)%; U = 0.000, P < 0.05], lower proportion of NK1.1+ DX5+ cells [(21.70 ± 12.48)% vs. (45.75 ± 2.26)%; U = 0.000, P < 0.05] and higher IFN-γ expression [(16.74 ± 1.36)% vs. (8.13 ± 1.90)%; U = 0.000, P < 0.05] were detected in the infected group than in the control group, while no significant difference was seen in TNF-α expression between the two groups [(67.98 ± 9.20)% vs. (52.93 ± 10.42)%; U = 2.000, P > 0.05]. The mouse uNK cells showed a strong cytotoxicity in the infected group, and the cytotoxicity gradually increased with the effector/target cell ratio. The cytotoxicity of uNK cells against YAC-1 cells was 2.30%, 4.32%, 8.12% and 12.65% in the infected group and 1.21%, 1.63%, 2.51% and 3.22% in the control group at effector/target cell ratios of 1:1, 5:1, 10:1 and 20:1, respectively. Following co-culture of human uterine lymphocytes and tachyzoites of the T. gondii RH strain for 48 h, the proportion [TOX 2:1 group vs. control group: (6.61 ± 1.75)% vs. (17.48 ± 4.81)%; F = 7.307, P < 0.01], and absolute number of human uNK cells in uterine lymphocytes of human uNK cells in uterine lymphocytes [TOX 2:1 group vs. control group: (12 104 ± 5 726) cells/well vs. (65 285 ± 21 810) cells/well; H = 11.540, P < 0.01] were significantly lower in the infected group than in the control group. A lower proportion of CD56brightCD16- NK cells [TOX 2:1 group vs. control group: (25.25 ± 5.90)% vs. (36.03 ± 4.51)%; F = 3.213, P > 0.05] and higher proportion of CD56dimCD16+ NK cells [TOX 2:1 group vs. control group: (11.15 ± 2.15)% vs. (7.09 ± 2.24)%; F = 2.992, P > 0.05] were detected in uNK cells in the infected group than in the control group, and the ratio of CD56brightCD16- cells/CD56dimCD16+ cells was significantly lower in the infected group than in the control group [TOX2:1 group vs. control group: (2.37 ± 0.92) vs. (5.58 ± 2.39); H = 8.228, P < 0.05]. In addition, the proportion of CD11b+CD27- cells in human uNK cells was significantly higher in the infected group than in the control group [TOX 2:1 group vs. control group: (30.28 ± 6.91)% vs. (17.48 ± 4.67)%; H = 6.556, P < 0.05], while no significant differences were found between the two groups in terms of IFN-γ [TOX 2:1 group vs. control group: (14.13 ± 1.28)% vs. (15.19 ± 1.64)%; F = 1.639, P > 0.05] or TNF-α expression [TOX 2:1 group vs. control group: (54.76 ± 10.02)% vs. (50.33 ± 3.67)%; F = 0.415, P > 0.05]. Human uNK cells presented a strong cytotoxicity in the infected group, and the cytotoxicity gradually increased with the effector/target cell ratio. The cytotoxicity of human uNK cells against K562 cells was 11.90%, 28.11%, 49.91% and 73.35% in the infected group and 12.21%, 21.63%, 33.51% and 48.22% in the control group at effector/target cell ratios of 1:1, 5:1, 10:1 and 20:1, respectively. CONCLUSIONS: T. gondii infection presents diverse effects on the differentiation and secretion ability of mouse and human uNK cells. However, T. gondii infection causes a reduction in the absolute number and enhances the cytotoxicity of both mouse and human uNK cells.

[Diagnostic value of combined application of GATA3, SOX10 and p16 in triple negative breast carcinomas].

Objective: To investigate the expression of GATA3, SOX10, and p16 in triple-negative breast cancer (TNBC) and analyze their significance and correlation with clinicopathology. Methods: The expressions of GATA3, SOX10 and p16 in 53 cases of TNBC and 50 cases of non-TNBC were detected by immunohistochemical staining. Results: GATA3 and SOX10 were positive in 58.5%(31/53) and 75.5%(40/53) of TNBC, respectively. The expression of SOX10 was significantly higher than that in non-TNBC (P<0.05). SOX10 was positive in 17 of the 22 cases that lacked GATA3 expression (77.3%). The expression of p16 was significantly higher in the TNBC, and the co-expression with SOX10 was significantly increased (P<0.05). The sensitivity, specificity, and AUC under the ROC curve of SOX10 were higher than those of GATA3. The sensitivity of SOX10 was higher than that of p16, but the specificity was lower than that of p16. The AUC of SOX10 was higher than that of p16. AUC of combined detection of GATA3 and SOX10, SOX10 and p16 were higher than that of each antibody alone (P<0.05). The expression of GATA3, SOX10, and p16 had no significant correlation with age, tumor size, and lymph node metastasis. The expression of SOX10 and p16 in grade 3 and basal-like TNBC increased significantly, and their co-expression increased. Conclusions: The expressions of SOX10 and p16 in TNBC are significantly increased. SOX10 is a reliable marker for the diagnosis of TNBC and a supplement to GATA3. Whether p16 is a marker related to the prognosis of TNBC remains to be further studied.

[Clinicopathological features and prognosis of gastrointestinal stromal tumor with PDGFRA-D842V mutation].

Objective: Platelet-derived growth factor alpha (PDGFRA) mutations are respectively rare in gastrointestinal stromal tumors (GIST). Most GIST with PDGFRA exon 18 mutations including D842V mutation are highly resistant to imatinib. The treatment of GIST harboring PDGFRA primary drug-resistant mutation is a major challenge. This article aims to investigate clinicopathologic features of GIST with PDGFRA-D842V mutation and the efficacy of comprehensive treatment, providing a reference for clinical practice. Methods: A retrospective cohort study was conducted to collect the clinicopathological and follow-up data of patients with GIST harboring PDGFRA mutation who were diagnosed and treated in the GIST Clinic of Renji Hospital from January 2005 to May 2020. According to the mutation site, the enrolled patients were divided into D842V mutation group and non-D842V mutation group. The differences of clinicopathologic characteristics between the two groups were compared. Furthermore, overall survival and prognostic factors were analyzed. Results: A total of 71 patients with PDGFRA-mutant GIST were included in this study, including 47 cases of D842V mutation (66.2%) and 24 cases of non-D842V mutation (33.8%). There were 28 male patients and 19 female patients in D842V mutation group, with a median age of 60 (36-82) years. There were 16 male patients and 8 female patients in non-D842V mutation group, with a median age of 62 (30-81) years. There were no significant differences in age, gender, primary location, surgical procedure, tumor size, mitotic count, expression of CD117 and DOG1, Ki-67 proliferation index and modified NIH grade between the two groups (all P>0.05). The positive rate of CD34 was 89.4% (42/47) and 62.5% (15/24) in the D842V mutation group and the non-D842V mutation group, respectively, with a statistically significant difference (χ(2)=5.644, P=0.018). Among all the cases, 66 cases underwent R0 resection without preoperative treatment; two cases underwent emergency operation with R1 resection because of tumor rupture; 2 cases were not operated after the pathological and mutation types were confirmed by biopsy (one case received avapritinib treatment and obtain partial remission). One case was diagnosed as wild-type GIST per needle biopsy in another institute, and underwent R0 resection after preoperative imatinib treatment for 6 months. After surgery, 5 high-risk GIST patients with D842V mutation and 5 high-risk GIST patients with non-D842V mutation were treated with imatinib for more than one year. The median follow-up time was 37 (1-153) months. As of the last follow-up among the patients who received R0 resection, 4 patients with D842V mutation had relapse, of whom 1 was in the period of imatinib administration, and the 3-year relapse-free survival rate was 94.2%; none of the patients with non-D842V mutation had relapse. There was no statistically significant difference in relapse-free surivval between two groups (P=0.233). Univariate analysis revealed that mitotic count (P=0.002), Ki-67 proliferation index (P<0.001) and modified NIH grade (P=0.025) were the factors associated with relapse-free survival of patients with D842V mutation after R0 resection (all P<0.05). However, the above factros were not testified as independant prognostic facors in multivariate Cox analysis (all P<0.05). Conclusion: Clinicopathologic features and the efficacy of radical resection in patients with PDGFRA-D842V mutation are similar to those in patients with non-D842V mutation.

Study on the function and mechanism of long non-coding RNA DMTF1v4 in the occurrence of colon cancer.

OBJECTIVE: To investigate the expression of long non-coding RNA (lncRNA) DMTF1v4 in colon cancer, and the relationship between its expression and disease occurrence. MATERIALS AND METHODS: Human colon cancer tissues and para-carcinoma tissues were harvested. The expression of lncRNA DMTF1v4 was measured by semi-quantitative PCR. The expression of DMTF1v4 in HT-29 colon cancer cells was downregulated using siRNA, and the effect of its downregulation on cell growth was determined by MTT assay and plate clone assay. The effect of DMTF1v4 downregulation on colon cancer cell migration was determined using a transwell assay and scratch wound assay. The effect of DMTF1v4 on colon cancer cell apoptosis was determined using Annexin V/PI double-staining. The changes in p-ERK, p-JNK, and p-p38 were measured by Western blot. HT-29 cells with downregulated DMTF1v4 expression were used to establish the subcutaneous heterotopic transplantation tumor model in nude mice to study the effect of DMTF1v4 on tumor growth in animals. RESULTS: Compared with para-carcinoma tissue, lncRNA DMTF1v4 in colon cancer tissue was highly expressed (p<0.001). Downregulating lncRNA DMTF1v4 in HT-29 cells showed that lncRNA DMTF1v4 promotes cell proliferation and migration, and suppresses apoptosis (p<0.05). The effect of lncRNA DMTF1v4 on the ERK/MAPK signaling pathway was evaluated. The expression of p-ERK, p-JNK, and p-p38 was increased significantly compared with the control group (p<0.01). The effect of downregulating DMTF1v4 on tumor growth in animals showed that tumor growth in nude mice was decreased, and the expression of apoptosis-related proteins was increased (p<0.01). CONCLUSIONS: The expression of lncRNA DMTF1v4 is elevated in colon cancer tissues; lncRNA DMTF1v4 promotes colon cancer cell proliferation and migration, and inhibits apoptosis by downregulating the expression of p-ERK, p-JNK, and p-p38, thus affecting the progression of colon cancer. This will provide a basis for the development of new clinical treatments for colon cancer.

Effects of sacral nerve stimulation with acupuncture on gut transit time and c-kit expression in colon of rats with slow transit constipation.

Sacral nerve stimulation (SNS) is an alternative surgical approach to alleviate fecal incontinence and constipation. This study aimed to explore the effects and underlying mechanisms of SNS with acupuncture on gut transit time and colon c-kit protein expression in rats with slow transit constipation (STC). Fifty Sprague-Dawley rats were randomly divided into five groups: blank control, SNS, Mosapride, sham SNS, and STC model control group. The STC model was established by subcutaneous injection of morphine. Each group was treated over a 15-day period. Gut transit time was measured 1 day before the treatment started and after 5, 10, and 15 days of treatment. After the 15-day treatment, animals were sacrificed and colonic tissues were collected for analysis of c-kit protein expression, using western blot analysis. We found significant differences in gut transit time in the SNS group compared with the Mosapride group after 5 (P = 0.001) and 10 (P = 0.004) days of treatment. After 15 days of treatment, there were no differences in gut transit time among the SNS, Mosapride, and blank control groups. However, significant differences were observed when comparing the SNS and Mosapride groups with the STC model and sham SNS groups. A decreased c-kit protein expression was observed in the STC model control, sham SNS, and Mosapride groups, compared with the SNS group (P = 0.001). Our data indicate that SNS can decrease gut transit time and increase the expression of c-kit protein in rats with STC to improve colon transit function.

Distribution of common pathogens in patients with pyogenic liver abscess in China: a meta-analysis.

Pyogenic liver abscess (PLA) is a potentially life-threatening disease in many parts of the world, especially in Asia. The aim of this study was to quantify the proportion of common pathogens in patients with PLA in China, using a meta-analysis method based on systematic review of published studies. Several electronic databases were searched to identify the studies reporting the pathogens of PLA. We performed a meta-analysis to calculate the pooled proportion of pathogens and subgroup analysis among the included studies using R 3.1.1 software. In total, 183 studies were included in our final analysis, Klebsiella spp (54 %), Escherichia spp (29 %), Enterobacter spp (9 %), Proteus spp (6 %) and Pseudomonas spp (5 %) comprised the major gram-negative bacteria. Gram-positive bacteria mainly included Staphylococcus spp (13 %), Streptococcus spp (8 %) and Enterococcus spp (7 %). The distribution of pathogens in PLA patients were different in different economic regions in China. The proportion of Klebsiella spp had an upward tendency in recent years compared to other pathogens. In addition, the proportion of common pathogens in PLA patients with diabetes mellitus (DM) were carried out indicating that the dominant pathogens were Klebsiella spp (66 %), Escherichia spp (21 %) and Enterobacter spp (11 %). This meta-analysis showed that the main pathogens of PLA were Klebsiella spp, Escherichia spp, Staphylococcus spp, and Enterobacter spp in China. To ensure a precise estimate of the epidemiology of the pathogens, further large-scale or even a population-based study is needed.

The taste-visual cross-modal Stroop effect: An event-related brain potential study.

Event-related potentials (ERPs) were recorded to explore, for the first time, the electrophysiological correlates of the taste-visual cross-modal Stroop effect. Eighteen healthy participants were presented with a taste stimulus and a food image, and asked to categorize the image as "sweet" or "sour" by pressing the relevant button as quickly as possible. Accurate categorization of the image was faster when it was presented with a congruent taste stimulus (e.g., sour taste/image of lemon) than with an incongruent one (e.g., sour taste/image of ice cream). ERP analyses revealed a negative difference component (ND430-620) between 430 and 620ms in the taste-visual cross-modal Stroop interference. Dipole source analysis of the difference wave (incongruent minus congruent) indicated that two generators localized in the prefrontal cortex and the parahippocampal gyrus contributed to this taste-visual cross-modal Stroop effect. This result suggests that the prefrontal cortex is associated with the process of conflict control in the taste-visual cross-modal Stroop effect. Also, we speculate that the parahippocampal gyrus is associated with the process of discordant information in the taste-visual cross-modal Stroop effect.

Limit on the electron neutrino magnetic moment from the kuo-sheng reactor neutrino experiment.

A search of neutrino magnetic moment was carried out at the Kuo-Sheng Nuclear Power Station at a distance of 28 m from the 2.9 GW reactor core. With a high purity germanium detector of mass 1.06 kg surrounded by scintillating NaI(Tl) and CsI(Tl) crystals as anti-Compton detectors, a detection threshold of 5 keV and a background level of 1 kg(-1) keV(-1) day(-1) at 12-60 keV were achieved. Based on 4712 and 1250 h of reactor ON and OFF data, respectively, the limit on the neutrino magnetic moment of mu(nu;(e))<1.3x10(-10)mu(B) at 90% confidence level was derived. An indirect bound of the nu;(e) radiative lifetime of m(3)(nu)tau(nu)>2.8x10(18) eV(3) s can be inferred.