Improving intensity-modulated radiation therapy quality assurance by adopting statistical process control.

PURPOSE: To use statistical process control for intensity-modulated radiation therapy (IMRT) quality assurance (QA) and improve tolerance limits and action limits. METHODS: An electronic portal imaging device (EPID) was selected to verify IMRT QA. The I-chart and the exponentially weighted moving averages (EWMA) chart were used to analyze the corresponding results. RESULTS: Twenty samples were used to enable the sampling requirements for building the control limits to be met. The I-chart showed that isolated data points beyond the control limits were mainly derived from complex plans. The EWMA made predictions of systematic errors earlier than the I-chart. Systematic errors primarily originated from the dose calibration on the EPID, and recalibrating the EPID could eliminate such errors. CONCLUSION: Statistical process control is an effective tool to detect controllable and can be used in IMRT QA. After calibrating the EPID, the tolerance and action limits all improved and satisfied the requirements/recommended values of the AAPM TG-218 report.

[Effects of urban construction on Oncomelania snails control in Wuhan City].

OBJECTIVE: To evaluate the effectiveness of urban construction projects on Oncomelania snails control in Wuhan City, so as to provide insights into the development of an integrated control strategy for urban schistosomiasis. METHODS: The data pertaining to the endemic situation of schistosomiasis, schistosomiasis control data, the construction of the Yangtze River Bridge, marshland management and park constructions were collected from Wuhan City from 1990 to 2020. The changes of areas of snail habitats and high-risk settings were compared before and after implementation of these urban construction projects to evaluate the effectiveness of urban construction projects on Oncomelania snails control. RESULTS: The number of schistosomiasis cases decreased by 97.35%, and the number of cattle with schistosomiasis decreased by 100% in Wuhan City from 1990 to 2020, with a 41.99% reduction in areas of snail habitats, including 94.97% and 34.40% reductions in snail habitats inside and outside the embankment decreased. During the period from 1990 through 2020, a total of 10 Yangtze River bridges were built in Wuhan City, and areas of snail habitats around the bridges reduced from 11 699.05 hm2 before the bridge building to 8 726.14 hm2 after the building (a 25.41% reduction), while the areas of high-risk settings reduced from 411.69 hm2 before the bridge building to 276.78 hm2 after the building (a 32.77% reduction). Following treatment of three marshlands in Hankou, Hanyang and Wuchang, the areas of snail habitats reduced from 225.80 hm2 before the management to 199.47 hm2 after the management (a 11.66% reduction), and the areas of high-risk settings reduced from 23.14 hm2 before the treatment to 17.73 hm2 after the building (a 23.38% reduction). Following the building of 5 parks in snail-infested settings, the areas of snail habitats reduced from 319.61 hm2 before the construction to 280.80 hm2 after the construction (a 12.14% reduction), and the areas of high-risk settings reduced from 35.00 hm2 before the construction to 27.73 hm2 after the construction (a 20.77% reduction). CONCLUSIONS: Simultaneous implementation of urban construction projects and schistosomiasis control measures is effective to shrink snail-infested setting and high-risk settings and control the endemic situation of schistosomiasis in endemic foci in Wuhan City.

[Epidemic trend and control progress of soil-transmitted nematodiasis in Jiangsu Province].

Soil-transmitted nematodiasis was once widely prevalent in Jiangsu Province, which seriously threatened human health and hindered socioeconomic development. The control efforts over decades resulted in a remarkable decline in the prevalence of soil-transmitted nematode human infections in Jiangsu Province, with a reduction from 59.32% in 1989 to 0.12% in 2019, and the human prevalence remains at < 0.5% since 2013. Since 1987, an integrated strategy has been adopted for the control of soil-transmitted nematodiasis in Jiangsu Province; however, the core interventions varies at different stages, which mainly include deworming, water and sanitation service improvement, health education, and monitoring and assessment. The criteria of effective soil-transmitted nematodiasis control had been achieved in all epidemic counties (districts) of Jiangsu Province by 2019. Further actions to strengthen health education and monitoring and implement precision control measures are required to consolidate the achievements of soil-transmitted nematodiasis control and eliminate the harm of soil-transmitted nematodiasis to humans. This review summarizes the epidemiology, control progress and evolution of control strategy of soil-transmitted nematodiasis in Jiangsu Province.

Long noncoding RNA ZFPM2-AS1 regulates ITGB1 by miR-1226-3p to promote cell proliferation and invasion in hepatocellular carcinoma.

OBJECTIVE: Long noncoding RNA (lncRNA) is emerging as a vital regulator in various tumors. However, the biological function of ZFPM2-antisense RNA 1 (ZFPM2-AS1) in hepatocellular carcinoma (HCC) remains unclear. The present study aims to explore the function and mechanism of ZFPM2-AS1 in hepatocellular carcinoma progression. PATIENTS AND METHODS: The ZFPM2-AS1 expression in HCC cells and tissues was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Effects of ZFPM2-AS1 on tumor cell proliferation and invasion were detected by CCK8 assay or EdU assay or matrigel migration assay and Western blot. The Luciferase reporter assay, RNA pulldown assay, qRT-PCR, and Western blot were performed to explore and confirm the interaction between ZFPM2-AS1 and miR-1226-3p and integrin ß1 (ITGB1). RESULTS: ZFPM2-AS1 was overexpressed in HCC tissues and cell lines. High levels of ZFPM2-AS1 were correlated with advanced TNM stage, distant metastasis and a poorer overall survival rate. ZFPM2-AS1 knockdown inhibited cell proliferation and invasion. ZFPM2-AS1 could directly bind to and negatively regulate miR-1226-3p expression. Moreover, ITGB1 was identified as a target gene of miR-1226-3p. ITGB1 was found to be directly negatively regulated by miR-1226-3p and indirectly upregulated by ZFPM2-AS1. Rescue assays demonstrated that ZFPM2-AS1 promotes HCC cell proliferation and invasion through modulating miR-1226/ITGB1 axis. CONCLUSIONS: ZFPM2-AS1 promotes cell proliferation and migration by regulating miR-1226-3p/ITGB1 axis in HCC.

[Investigation on the risk of of Anisakis infection among high - risk populations along the coastal areas of Jiangsu Province].

OBJECTIVE: To investigate the risk of Anisakis infections among high-risk populations along the coastal areas of Jiangsu Province, so as to develop the strategy for the prevention and control of anisakiasis in the province. METHODS: Three counties along the coastal areas of Jiangsu Province were selected as the study sites in 2018, including Rudong County in Nantong City, Haizhou District in Lianyungang City and Dongtai City in Yancheng City. The knowledge, attitude and practice (KAP) of anisakiasis prevention and control, and the prevalence of serum specific IgG antibody against Anisakis were investigated among high-risk populations among these three study sites, including fishermen, fish seller and people who liked eating fresh and live marine fish. Factors affecting the prevalence of the specific IgG antibody against Anisakis were identified using a multiple logistic regression model. In addition, Anisakis larvae infections were detected in fresh and live marine fish samples collected from local markets, and the prevalence and intensity of Anisakis infections were estimated. RESULTS: A total of 625 high-risk populations were investigated, including 349 men (55.8%). Only 13.0% of the subjects heard about anisakiasis, and a low awareness rate of anisakiasis prevention and control knowledge was seen among these three types of high-risk populations. There were 21.6% of the subjects eating raw or half-cooked marine fish, 5.8% eating undercooked marine fish, 3.2% presenting vomiting, nausea and diarrhea after eating marine fish, 5.1% developing systemic allergic symptoms, and 65.6% using the same chopping board for raw and cooked food. The sero-prevalence of the anti-Anisakis IgG antibody was 7.0% among the study subjects. Multiple logistic regression analysis identified education level [OR = 0.687, 95% CI (0.478, 0.987)] and development of systemic allergic symptoms [OR = 4.641, 95% CI(1.411, 15.268)]as factors affecting the positive anti-Anisakis IgG antibody among the study subjects. Among 494 fresh and live marine fish detected, the prevalence and intensity of Anisakis larvae infection was 64.0% and 8.1 larvae per fish, with high prevalence seen in Trichiurus haumela and Pneumatophorus japonicas. CONCLUSIONS: The awareness of anisakiasis prevention and control knowledge is low among the high-risk populations living along the coastal areas of Jiangsu Province, and there are high-risk behaviors, such as eating raw or half-cooked food, using the same chopping board for raw and cooked food. In addition, the prevalence of Anisakis infections is high in the marine fish in these areas. Therefore, the health education and health promotion for anisakiasis prevention and control should be intensified.

[Study on the expression of FoxM1 in chronic rhinosinusitis and the effect of inflammatory factors on FoxM1 expression].

Objective: To check Forkhead box M1 (FoxM1) expression in nasal mucosal of chronic rhinosinusitis (CRS) patients and the effect of inflammatory factors on FoxM1 expression, in order to research the significance of FoxM1 in CRS. Methods: From January to October of 2018, 50 patients hospitalized in the Department of Otorhinolaryngology, Head and Neck Surgery, First Affiliated Hospital of Nanchang University were enrolled in this study. Twenty CRS patients with polyps (CRSwNP), 20 CRS patients without nasal polyps (CRSsNP) and 10 patients with simple deviation of nasal septum (the control groups) were selected. The expression of FoxM1 in nasal mucosa of these patients was detected by immunohistochemistry (IHC) and real-time fluorescent quantitative PCR (qRT-PCR). Meanwhile, HE stain was used to observe the pathologic changes in each sample. By establishing human nasal epithelium cells cultivating model in vitro and identifying via immumofluorescence method, experimental group and control group were set up, then activation factors including interleukin (IL)-1ß, IL-4, IL-5, IL-17, interferon-γ (IFN-γ) and staphylococcal entemtoxin B (SEB) were added in the models after stabilizing passage, and qRT-PRC and Western blot method were applied to check the expressing change of FoxM1. Software SPSS 18.0 was used for statistical analysis. Results: HE stain showed that the mainly pathologic change in nasal mucosa of CRS patients with or without nasal polyp was mucosal epithelial cells, goblet cell and submucosal gland hyperplasia, accompanied by a large number of inflammatory cells infiltration. The result of IHC demonstrated that both of the expression of FoxM1 in nasal mucosal tissue of CRS patients in the CRSwNP and CRSsNP groups exceed that of the control group (80% vs 75% vs 20%, χ(2) value was 10.000, 8.213, respectively, all P<0.05); there was no difference of expression between the two groups of CRS patients (χ(2)=0.143, P>0.05). The result of qRT-PCR demonstrated that the expression of FoxM1 mRNA in nasal mucosa of CRSwNP and CRSsNP was increased compared with that of the control group (3.309±1.511 vs 3.261±1.336 vs 1.000±0.774, t value was 4.519, 4.928, respectively, all P<0.05), but the difference between the two groups of CRS patients had no statistic significance (t=0.107, P=0.909). Nasal mucosa epithelial cells cultivating models was established successfully. Q-RT PCR and Western blot were conducted after stimulation of 100 ng/ml IL-1ß, IL-4, IL-5, IL-17, IFN-γ and SEB for 36 h, and the proteins expression levels of FoxM1 exceeded the groups without stimulation with statistic significance. Conclusions: The expression of FoxM1 in CRS increases and many types of cytokine can induce the increase of FoxM1 in human nasal epithelial cells. FoxM1 may participate in the process of pathogenesis in CRS.

[Recurrent maxillary sinus pleomorphic gonadoma into myoepithelial carcinoma: a case report].

Summary A 44-year-old woman with left nasal obstruction and facial numbness for 4 months was admitted to hospital. The patient did not have amblyopia, vision loss, runny nose with blood, dizziness ,headache or other discomfort.In 1991 and 2001, the patient were pathologically diagnosed as pleomorphic adenomas.CT of nasal cavity and paranasal sinuses showed that in the left maxillary sinus there was an about 4.4 cm×4.5 cm×4.7 cm large mass soft tissue density, showing expansive growth protruding into the left orbital floor.MRI showed that the lumped short T1 signal was seen in the left maxillary sinus and the linear long T1 signal was seen in the left nasal cavity, and the liquid accumulation signal foci could be seen in the left maxillary sinus.Postoperative pathological findings: （left maxillary sinus mass） Combining morphology, immunohistochemical results and medical history, consistent with pleomorphic adenoma carcinogenesis （cancer in pleomorphic adenoma）, carcinogenesis type is myoepithelial carcinoma.

Overexpression of long non-coding RNA MINCR contributes to progressive clinicopathological features and poor prognosis of human hepatocellular carcinoma.

OBJECTIVE: MYC-induced long non-coding RNA (MINCR) has been shown to be a long noncoding RNA that facilitates the progression of a number of malignancies, including hepatocellular carcinoma (HCC). However, few studies have explored the expression and role of MINCR in HCC. In this study, we aimed to investigate the clinical significance of MINCR in HCC. PATIENTS AND METHODS: MINCR expression levels in 161 pairs of HCC tissues and pair-matched adjacent normal tissues were examined by qRT-PCR. The correlation between clinicopathological features and MINCR expression was analyzed by x2 test. Differences in patient survival were determined using the Kaplan-Meier method and a log-rank test. The significance of survival variables was analyzed using the Cox multivariate proportional hazards model. RESULTS: Our results showed that MINCR was significantly upregulated in HCC tissues, compared with paired adjacent nontumor tissue samples. MINCR upregulation was correlated with TNM stage (p = 0.005) and histological grade (p = 0.001). The results of Kaplan-Meier method and log-rank test indicated that the 5-years overall survival of the high MINCR group was significantly lower than that of low MINCR group (p = 0.0035). Univariate and multivariate analysis results indicated that MINCR was an independent prognostic factor in HCC (p < 0.05). CONCLUSIONS: We firstly provided the possibility that evaluating MINCR in HCC tissues may have prognostic and predictive value in the clinical management of HCC patients.

[Value of double contrast-enhanced ultrasound QontraXt three-dimensional pseudocolor quantitative analysis to therapeutic effect evaluation of preoperative neoadjuvant chemotherapy in advanced gastric cancer patients].

Objective: To investigate the value of tumor perfusion parameter measured by using double contrast-enhanced ultrasound (DCEUS) QontraXt three-dimensional pseudocolor quantitative analysis to the therapeutic effect evaluation of preoperative neoadjuvant chemotherapy (NAC) in advanced gastric cancer (AGC) patients. Methods: Eighty-nine AGC patients underwent 3 cycles of preoperative NAC (XELOX) followed by complete resection of lesion. The DCEUS QontraXt three-dimensional pseudocolor was performed one or two weeks before the NAC and operation were applied, respectively. The peak enhancement (PE), time to peak (TP), sharpness of the bolus (ß) and area under the enhancement curve (AUC) of primary gastric tumor were measured by QontraXt three-dimensional pseudocolor quantitative analysis. These DCEUS parameters between respond and non-respond groups before and after NAC therapy were compared. The prediction accuracy of DCEUS to the therapeutic effect evaluation of preoperative NAC was determined by the receive operating characteristic (ROC) curves. Results: Among 89 AGC patients, 52 patients responded to NAC therapy, while 37 patients resisted to NAC therapy. Twelve cases in respond group and 26 cases in non-respond group were mucinous carcinoma. Forty cases in respond group and 11 cases in non-respond group were non-mucinous carcinoma (P<0.05). In responder group, the PE and TP before NAC were (53.7±9.3)% and (14 521±2 667) ms, and (32.2±5.5)% and (17 235±1 898) ms after NAC. The ratio of changes of PE (ΔPE) and TP (ΔTP) were 0.43±0.17 and 0.36±0.14, respectively. In non-respond group, the PE and TP before NAC were (54.4±7.2)% and (13 869±3 247) ms, and (45.3±6.1)% and (15 127±1 423) ms after NAC therapy. The ratio of ΔPE and ΔTP were 0.24±0.20 and 0.22±0.12. The PE and TP after NAC, the ratio of ΔPE and ΔTP were significant different among these two groups (all of P<0.05). The ROC curves showed that the ratio of ΔPE in assessing the respond of gastric cancer patients to NAC was superior compared to other parameters (AUC=0.784, P=0.004). The optimal cut-off value of the ratio of ΔPE was 24% and its sensitivity and specificity to the therapeutic effect evaluation of NAC in gastric cancer were 82.7% and 64.9%. Conclusion: DCEUS QontraXt three-dimensional pseudocolor quantitative analysis might be a novel, noninvasive and reliable method to evaluate the therapeutic effect of preoperative NAC in AGC patients.

[Clinical analysis of 42 cases of nasal natural killer/T-cell lymphoma prominently presenting as ocular symptoms].

Objective:To investigate the clinical features and prognosis of nasal natural killer/T-cell lymphoma(NKTL) with obvious ocular symptoms.Method:We retrospectively analyzed the clinical data of 42 patients with nasal NKTL who prominently showed eye symptoms and were treated from January 2011 to December 2017.Result:After treatment,the patients received complete remission(CR) in 19 cases and partial remission(PR) in 9 cases. The total effective rate was 66.67%.Median followup time was 39 months and 18 patients died.In this study, there was no correlation between sex and prognosis(P>0.05).Age＞60 years was not related to the prognosis of the patients(P>0.05).The 3-year survival rate of patients treated with radiotherapy and chemotherapy was higher than that of patients treated with radiotherapy or chemotherapy alone(73.3%, 16.7%，P<0.01). The 3-year survival rate of patients with B symptoms was 41.7%, which was not related to prognosis(P>0.05).The prognosis of EBV-DNA positive patients was poor(P<0.01). Twenty-six patients with stage Ⅰ and Ⅱ had a better prognosis(P<0.01). Patients with international prognostic index(IPI) ≤1 had a better prognosis than those with IPI≥2(83.3 %, 37.5%，P<0.01).Conclusion:Nasal NKTL lymphomas with prominent ocular symptoms are difficult to diagnose early and easy to be misdiagnosed. Clinicians should make a definite diagnosis by pathological biopsy and immunohistochemistry as soon as possible, and assist in diagnosis by EBER in situ hybridization if necessary.

Histone 3 lysine 9 acetylation is a biomarker of the effects of culture on zygotes.

Acetylation of histone proteins is a major determinant of chromatin structure and function. Fertilisation triggers a round of chromatin remodelling that prepares the genome for the first round of transcription from the new embryonic genome. In this study we confirm that fertilisation leads to a marked progressive increase in the level of histone 3 lysine 9 acetylation in both the paternally and maternally derived genomes. The culture of zygotes in simple defined media caused a marked increase in the global level of acetylation and this affected the male pronucleus more than the female. The culture created a marked asymmetry in staining between the two pronuclei that was not readily detected in zygotes collected directly from the reproductive tract and was ameliorated to some extent by optimized culture media. The increased acetylation caused by culture resulted in increased transcription of Hspa1b, a marker of embryonic genome activation. Pharmacological analyses showed the hyperacetylation of H3K9 and the increased expression of Hspa1b caused by culture were due to the altered net activity of a range of histone acetylases and deacetylases. The marked hyperacetylation of histone 3 lysine 9 caused by culture of zygotes may serve as an early biomarker for the effects of culture on the normal function of the embryo. The results also provide further evidence for an effect of the stresses associated with assisted reproductive technologies on the normal patterns of epigenetic reprogramming in the early embryo.

The induction of tumour suppressor protein P53 limits the entry of cells into the pluripotent inner cell mass lineage in the mouse embryo.

The early preimplantation embryo is susceptible to a range of exogenous stresses which result in their reduced long-term developmental potential. The P53 tumour suppressor protein is normally held at low levels in the preimplantation embryo and we show that culture stress induces the expression of a range of canonical P53-response genes (Mdm2, Bax and Cdkn1a). Culture stress caused a P53-dependent loss of cells from resulting blastocysts, and this was most evident within the inner cell mass population. Culture stress increased the proportion of cells expressing active caspase-3 and undergoing apoptosis, while inhibition of caspase-3 increased the number of cells within the inner cell mass. The P53-dependent loss of cells from the inner cell mass was accompanied by a loss of NANOG-positive epiblast progenitors. Pharmacological activation of P53 by the MDM2 inhibitor, Nutlin-3, also caused increased P53-dependent transcription and the loss of cells from the inner cell mass. This loss of cells could be ameliorated by simultaneous treatment with the P53 inhibitor, Pifithrin-α. Culture stress causes reduced signalling via the phosphatidylinositol-3-kinase signalling pathway, and blocking this pathway caused P53-dependent loss of cells from the inner cell mass. These results point to P53 acting to limit the accumulation and survival of cells within the pluripotent lineage of the blastocyst and provide a molecular framework for the further investigation of the factors determining the effects of stressors on the embryo's developmental potential.

Surgical treatment of incarcerated calculi via laparoscopic bile duct exploration using laparotomy biliary lithotomy forceps.

The present study aimed to investigate the practicability and clinical value of applying laparotomy biliary lithotomy forceps to laparoscopic bile duct exploration (LCBDE) for the surgical treatment of incarcerated calculi. A total of 63 patients were diagnosed with cholecystolithiasis and choledocholithiasis. The present study performed a retrospective analysis of clinical samples from 16 of these patients who had incarcerated calculi at the terminus of the common bile duct, and who had been treated with laparoscopic cholecystectomy and LCBDE. During the procedure, laparotomy biliary lithotomy forceps were used to gently remove the calculi from the common bile duct. Of the surgical procedures that used laparotomy biliary lithotomy forceps, one case was unsuccessful and 15 cases were successful. The results of the present study suggested that it may be clinically advisable to use laparotomy biliary lithotomy forceps to remove incarcerated calculi from the common bile duct during a laparoscopy, since it is easy, economical and effective.

Nuclear factor-like factor 2-antioxidant response element signaling activation by tert-butylhydroquinone attenuates acute heat stress in bovine mammary epithelial cells.

Nuclear factor (erythroid-derived 2)-like factor 2 (Nrf2) is a transcription factor that binds to the antioxidant response element (ARE) in the upstream promoter region of many antioxidative genes. The Nrf2-ARE signaling plays a key role in the cellular antioxidant-defense system, but whether Nrf2 activation has protective effects against heat shock (HS) stress in mammary epithelial cells (MEC) remains unclear. The objective of this study was to determine whether tert-butylhydroquinone (tBHQ), a well-known Nrf2 activator, could attenuate heat stress-induced cell damage in MAC-T cells of the bovine MEC line. The MAC-T cells were exposed to HS (42.5°C for 1h) followed by recovery at 37°C to mimic HS. Compared with cells that were consistently cultured at normothermia (37°C), the cell viability levels significantly decreased after HS stress. In parallel, heat stress increased the reactive oxygen species levels and induced cellular apoptosis and endoplasmic reticulum stress. The MAC-T cells that were pretreated with tBHQ (10µM) for 2h followed by HS had a reduction in the loss of cell viability. The tBHQ pretreatment significantly decreased cellular reactive oxygen species levels and stress-related marker gene expression. The tBHQ-treated MAC-T cells showed strong Nrf2-ARE signaling activation and a nuclear accumulation of Nrf2 and upregulated expression of Nrf2-ARE downstream genes. Small interfering RNA silencing of Nrf2 in HS-treated MAC-T cells almost completely abolished the cytoprotective effects by tBHQ. Overall, our results demonstrated that HS could cause cell damage in cultured bovine MEC, and that activation of Nrf2 by tBHQ could attenuate HS-induced cell damage.

[Expression of Foxm1 in chronic sinusitis and itsrelationship with MUC5AC].

Objective:To explore the expression of Forkhead box protein m1(Foxml) in the sinus mucosa of patients with chronic rhinosinusitis(CRS) and the relationship between Foxm1 and MUC5AC for further understanding of the pathogenesis of CRS.Method:We obtained the sinus mucosa from 25 CRS with polyps(CRSwNP) patients and 25 CRS without polyps (CRSsNP) patients as two experimental groups.Nasal mucosa from 15 normol cases were obtained as control group.We used HE,Periodic acid-schiff staining to observe the histopathological change of each sample.The expression of Foxm1 and MUC5AC were determined by immunohistochemistry(IHC) and qRT-PCR in each group.We also analysed the relationship between Foxm1 and MUC5AC.Result:The HE and PAS staining showed that in the CRSwNP and CRSsNP patients,the main histopathological features was the hyperplasia of goblet cell,inflammatory cell and submucosal gland.IHC showed that Foxm1 and MUC5AC expression were higher in CRSwNP and CRSsNP compared with normal sinus mucosa.qRT-PCR also revealed that Foxm1 and MUC5AC expression was higher in CRSwNP and CRSsNP than in normal mucosa(P<0.05).Foxm1 and MUC5AC had a significant positive correlation.Conclusion:The expressions of Foxm1 and MUC5AC were increased in CRS,and Foxm1 may play an important role in the process of hypersecretion of MUC5AC in CRS.

Survival signalling in the preimplantation embryo.

The development of the preimplantation embryo (from fertilisation until the formation of the differentiated blastocyst) occurs without a requirement for exogenous mitogenic or survival signals. This distinguishes the behaviour of cells in the early embryo from all other normal cells. The discovery that fertilisation triggers the production and release of potent bioactive mediators by the embryo that act back on membrane receptors demonstrated the presence of closed autocrine embryotrophic loops. It is now clear that these ligands act in concert with paracrine mediators normally present within the reproductive tract to support the normal development of the embryo. These ligands act via receptors expressed by the embryo to activate signalling transduced by 1-o-phosphatidylinositol-3-kinase and the resultant formation of phosphatidylinositol-3,4,5-trisphosphate. This polyphosphorylated membrane phospholipid acts as a docking site for proteins possessing the PH domain. These include PDK1, AKT and phospholipase C. The activation of these proteins accounts for the initiation of new transcription from the embryonic genome to form a pro-survival, anti-apoptotic transcriptome and the post-transcriptional activation of pro-survival signalling within embryonic cells. This includes the attenuation of action of pro-apoptotic signals, such as P53. The production of embryotrophic ligands after fertilisation bootstraps development by the activation of transcription from the embryonic genome, followed by the activation of pro-survival settings within embryo cells.

Physiological and morphological responses induced by α-particle radiation on Arabidopsis thaliana embryos.

Alpha (α)-particle radiation has been thoroughly studied in the occupational and residential environments, but biological mechanisms induced by α-particle radiation on plants are not clearly understood. In this study, radiation effects were examined using different total doses (1, 10, 100 Gy, respectively) of 241Am, α-particle on Arabidopsis embryos. No significant difference in the germination percentage was observed between the 3 levels of doses and the control. Germination speed and root length were increased by treatment with the 1-Gy dose of a-particles, and decreased by treatment with 10- and 100-Gy doses. Moreover, the bending degree of roots increased with radiation dose, and the roots showed an "S" shape when treated with the 100-Gy dose. Root bending under the 100-Gy dose was inhibited by scavengers of reactive oxygen species (ROS). Root gravitropism and root length may respond to the consistency of ROS induced by irradiation. Further analysis of the physiological effects revealed that an increase in a-particle radiation intensity enhanced the activity of catalase and the content of malondialdehyde, but superoxide dismutase activity was reduced by treatment with 100-Gy radiation of a-particles, suggesting that the high linear energy transfer of a-particles may cause a relatively high level of membrane lipid preoxidation and high accumulation of ROS. ROS showed both physiological and morphological responses following exposure to α-particle radiation in Arabidopsis embryos.

The regulation of the expression and activation of the essential ATF1 transcription factor in the mouse preimplantation embryo.

The co-expression of the CREB and ATF1 transcription factors is required for the development of preimplantation embryos. Embryotropin-mediated, calcium/calmodulin-dependent signalling activates CREB-induced transcription in the two-cell embryo, but the regulation of ATF1 in the embryo is not known. This study demonstrates that ATF1 begins to accumulate within both pronuclei of the mouse zygote by 20 h post-human chorionic gonadotrophin. This did not require new transcription (not blocked by α-amanitin), but was dependent upon protein synthesis (blocked by puromycin) and the activity of P38 MAP kinase. ATF1 becomes an active transcription factor upon being phosphorylated. A marked accumulation of phosphorylated ATF1 was evident in two-cell embryos and this persisted in subsequent stages of development. This phosphorylation was enhanced by the actions of autocrine embryotropic mediators (including Paf) and required the mutual actions of P38 MAP kinase and calmodulin-dependent pathways for maximum levels of phosphorylation. The combined inhibition of these two pathways blocked embryonic genome activation (EGA) and caused embryos to enter a developmental block at the two-cell stage. The members of the CREB family of transcription factors can generate one of the most diverse transcriptomes of any transcription factor. The demonstration of the presence of activated CREB and ATF1 within the embryonic nucleus at the time of EGA places these transcription factors as priority targets as key regulators of EGA.

Gpr97 is essential for the follicular versus marginal zone B-lymphocyte fate decision.

Gpr97 is an orphan adhesion GPCR and is highly conserved among species. Up to now, its physiological function remains largely unknown. Here, we show that Gpr97 deficiency results in an extensive reduction in B220(+) lymphocytes in mice. More intensive analyses reveal an expanded marginal zone but a decreased follicular B-cell population in Gpr97(-/-)spleen, which displays disorganized architecture characterized by diffuse, irregular B-cell areas and the absence of discrete perifollicular marginal and mantle zones. In vivo functional studies reveal that the mutant mice could generate antibody responses to T cell-dependent and independent antigens, albeit enhanced response to the former and weakened response to the latter. By screening for the molecular events involved in the observed phenotypes, we found that lambda 5 expression is downregulated and its upstream inhibitor Aiolos is increased in the spleen of mutant mice, accompanied by significantly enhanced phosphorylation and nuclear translocation of cAMP response element-binding protein. Interestingly, increased constitutive Nf-κb p50/p65 expression and activity were observed in Gpr97(-/-) spleen, implicating a crucial role of Gpr97 in regulating Nf-κb activity. These findings uncover a novel biological function of Gpr97 in regulating B-cell development, implying Gpr97 as a potential therapeutic target for treatment of immunological disorders.