The functional and regulatory entities underlying free and peptide-bound amino acid transporters in the bovine mammary gland.

Free and peptide-bound AA act as building blocks and key regulators of milk protein. To improve milk protein production, mammary epithelial cells of lactating mammals require extensive AA movement across the plasma membrane via multiple transport systems. Recent studies on bovine mammary cells/tissues have expanded the number of AA transporter systems identified and the knowledge on their contribution to AA utilization for milk protein synthesis and the regulatory machinery. However, in lactating cows, the exact intracellular location of mammary AA transporters and the extent of mammary net AA utilization for milk protein production remain unclear. This review highlights the existing knowledge on various characteristics, such as substrate specificity, kinetics, their effects on AA uptake and utilization, and regulatory mechanism, of recently examined bovine mammary free and peptide-bound AA transporters.

[Clinical and endoscopic characteristics of gastrointestinal mantle cell lymphoma].

Objective: To study the clinical and endoscopic features of mantle cell lymphoma (MCL). Methods: The clinical data and endoscopic characteristics of 20 patients with gastrointestinal MCL treated in the Affiliated Hospital of Zunyi Medical University and the Digestive Endoscopy Center of Guizhou Medical University from January 2011 to January 2021 were collected, and the histological and immunohistochemical characteristics of the tumor were observed. Meanwhile, 44 cases of non-gastrointestinal MCL in the above two hospitals in the same period were also enrolled. The clinical and endoscopic differences between the two groups were compared. Results: Among the 20 patients with gastrointestinal MCL, the male to female ratio was 4∶1 (16/4), the average age was (60.3±9.7) years, and 11 patients (55.0%, 11/20) were over 60 years old. The main clinical manifestations of 20 patients at the onset of the disease were bloody stool or black stool in 9 cases (45.0%), abdominal pain in 8 cases (40.0%). Moreover, 5.0% (1/20) had B symptoms (fever, night sweat and weight loss). The most frequently involved site was colorectum (50.0%). Endoscopic findings showed mucosal polypoid lesions in 13 cases (65.0%, 13/20), involving multiple intestinal segments, with polyp diameter of 0.2-3.5 cm. Two cases (10%) were in stage Ⅲ and 18 cases (90%) were in stage Ⅳ according to Ann Arbor staging. Seven cases (35.0%, 7/20) had the involvement of the peritoneal and retroperitoneal lymph nodes, 9 cases (45.0%, 9/20) had the involvement of the superficial lymph nodes in 45.0% (9/20), and 4 cases (20%, 4/20) had other extranodal invasion. Splenomegaly accounted for 35.0% (7/20). CD20, Bcl-2 and CD5 were positive or strongly positive in all gastrointestinal MCL tumor cells in 20 cases. Cyclin D1 was positive or strongly positive, accounting for 90.0% (18/20). The sex-determining region of Y chromosome-related high-mobility-group box 11 (SOX11) was positive or strongly positive, accounting for 65.0% (13/20). The positive index of Ki-67 was 5%-80%. Twenty cases (100%, 20/20) were followed up. The median follow-up was 41.2 months (1-74 months). There was no significant difference in the 3-year overall survival rate between the gastrointestinal MCL patients (70.0%) and non-gastrointestinal MCL patients (72.7%) (χ2=0.051, P=0.822). Conclusions: Abdominal discomfort is the main symptom of gastrointestinal MCL, which often occurs in the colorectum and often presents with multiple polypoid lesions. There are relatively few cases with B symptoms and extranodal invasion, and the prognosis is not different from that of patients with non-gastrointestinal MCL.

Exploring the nursing effect of application Albizia bark on autism in children based on network pharmacology and molecular docking.

OBJECTIVE: Autism is a disorder that manifests itself in early childhood. Early diagnosis of autism may not only help the affected children themselves, but also affect family well-being and social stability. The natural drug Albizia bark has been reported to have some effect in the prevention and treatment of autism in children. Therefore, we used network pharmacology and molecular docking to explore the possible mechanism. MATERIALS AND METHODS: TCMID and BATMAN-TCM was used to retrieve the chemical constituents of Albizia bark, and then obtained the relevant targets about autism by TTD, Gene Cards and OMIM. The resulting ingredients and targets were predicted, then a protein interaction network was constructed, and finally bioinformatics analysis was performed. Finally, molecular docking was used to verify the effective ingredients and targets obtained from the screening. RESULTS: Leucaena saponin B, luteolin, 3', 4', 7-trihydroxyflavone, which may be the key compounds for the treatment of autism. BP mainly involving signal transduction, G protein coupled receptor signal pathway, protein phosphorylation. CC, mainly involving plasma membrane, integral component of plasma membrane, MF, including protein binding, adenosine triphosphate binding, protein kinase activity. Molecular docking showed that AKT1, HRAS, PIK3CA, PIK3R1 and SRC, five potential targets, had good binding ability to Leucaena saponin B. CONCLUSIONS: The natural drug Albizia bark exerts pharmacological effects in a multi-component, multi-target and multi-channel manner, including neural regulation, inflammatory response and immune regulation.

[Analysis of gene mutation profile of adult soft tissue sarcomas using high-throughput sequencing technology].

Objective: To understand the genetic variation of soft tissue sarcomas, and to provide a scientific evidence for the individualized treatment. Methods: The somatic mutation and germline mutation of 45 adult soft tissue sarcomas had been detected by high-throughput sequencing technology, the clinical data were also analyzed. Results: A total of 88 gene mutations were detected in 45 samples, including 78 single nucleotide variation (SNV), 13 insertion/deletion (Indel) and 19 copy number variation (CNV). The most common mutant genes are TP53, CDKN2A, MDM2, CDK4, NF1 and PTEN. Among them, the mutation rates of TP53-MDM2/MDM4-CDKN2A pathway, CDKN2A/CDK4/RB1 pathway, and RAS/NF1/PTEN/PI3K pathway were more frequent (32/88, 36%). In terms of immunotherapy biomarkers among 10 samples, the median value of tumor mutation burden was 2.02 muts/Mb (0-4.24 muts/Mb), and all were microsatellite stable. Conclusions: This study analyzes the genetic variation of soft tissue sarcoma, and determines the high-frequency gene mutations and pathways, which may be the potential drug targets. This finding can provide scientific evidences for the personalized treatment of soft tissue sarcoma.

Effects of sarcopenia, hypoalbuminemia, and laparoscopic surgery on postoperative complications in elderly patients with colorectal cancer: A prospective study.

With the increasing number of elderly patients, the risk of diseases such as colorectal cancer (CRC) has increased. The objective of this prospective study was to explore the effects of sarcopenia, hypoalbuminemia, and laparoscopic surgery on postoperative complications among elderly patients who recently underwent colorectal surgery. Patients aged over 65 years who underwent surgery for CRC at the First Affiliated Hospital of Wenzhou Medical University were considered for this study. The demographical and clinical characteristics of these patients, as well as postoperative complications, were prospectively analyzed. The patients were divided into two groups depending on the diagnosis of sarcopenia, and the clinical variables corresponding to the two groups were compared. Further, the risk factors associated with postoperative complications were evaluated using univariate analysis and multivariate logistic regression analysis. A total of 360 patients fulfilled the inclusion criteria. Incidences of postoperative complications in the sarcopenia and non-sarcopenia groups were at 38.3% and 27.3%, respectively. In addition, sarcopenia (p=0.029) and hypoalbuminemia (p=0.010) were identified as independent risk factors, while laparoscopic surgery (p=0.023) was identified as a protective factor for postoperative complications. However, laparoscopic surgery was a protective factor for postoperative complications in the colon group only (p=0.001). Sarcopenia and hypoalbuminemia are independent risk factors that influence the probability of developing complications following CRC surgery. Laparoscopic surgery is a protective factor for postoperative complications of CRC patients, particularly colon cancer patients.

Effects of jugular infusions of isoleucine, leucine, methionine, threonine, and other amino acids on insulin and glucagon concentrations, mammalian target of rapamycin (mTOR) signaling, and lactational performance in goats.

The supply and profile of absorbed AA may affect milk protein synthesis through hormonal changes and mammalian target of rapamycin (mTOR) signaling pathways; and Ile, Leu, Met, and Thr (ILMT) are the 4 AA that have been reported to have the greatest effect on mammary mTOR signaling. The extent to which ILMT and the other remaining AA (RAA) differ in their effects on milk protein synthesis needs to be systematically investigated. In this study, 5 lactating goats, averaging 120 ± 10 d in milk, fitted with jugular vein and carotid artery catheters, were fasted for 24 h, followed by intravenous infusions of a mixture containing AA and glucose for 8 h in a 5 × 5 Latin square design. The AA mixtures were formulated according to the profile of casein. The amounts of AA infused were calculated based on supplies of AA when metabolizable protein (MP) was at requirement (MR). Treatments were an infusate containing glucose without AA (NTAA); an infusate containing 3 × the MR of Ile, Leu, Met and Thr (3F0R); and infusates containing 3F0R plus 1, 2, or 3 × MR of RAA (3F1R, 3F2R, and 3F3R, respectively) according to amounts provided when fed to meet MP requirements for maintenance and lactation for each goat. Milk, arterial blood, and mammary tissue samples were collected immediately after halting the infusion. Relative to NTAA, supplementation of ILMT tended to increase milk protein production and plasma glucose concentrations, and increased milk and lactose production, but had no effects on production or content of milk fat. Graded supplementation of RAA tended to quadratically affect production of milk and lactose. Arterial glucose and glucagon concentrations decreased linearly, and plasma insulin concentrations decreased quadratically with increased RAA. Mammary p70-S6K1 phosphorylation was decreased by addition of ILMT compared with NTAA but increased linearly with increased RAA infusion. Furthermore, EIF4EBP1 gene expression was much lower for 3F-treated goats than for the NTAA treatment. Both MTOR and RPS6KB1 gene expressions were decreased quadratically with increased RAA supply. These results suggested that short-term milk protein yield tended to be increased by elevated ILMT availability, and this trend was not explained by variations in mammary mTOR signaling or pancreatic hormone secretions, whereas graded increase of RAA in combination with ILMT appeared to regulate the efficiency of conversion of glucose to lactose in a manner not involving milk protein production.

[Treatment and analysis of prognostic factors of stage â ¡ and â ¢ undifferentiated high-grade pleomorphic sarcoma in extremities and trunk].

Objective: To evaluate the efficacy and prognostic factors of comprehensive treatment of undifferentiated high grade pleomorphic sarcoma (UHGPS) in extremities and trunk, including surgery, radiotherapy and chemotherapy. Methods: A retrospective analysis and follow-up of 131 UHGPS cases with clinical stage Ⅱ or Ⅲ in extremities and trunk soft tissue was performed to analyze the prognostic factors. Survival data were collected through follow-up. The survival rate was calculated with life table method and Kaplan-Meier survival curves were drawn. Survival rate between the two groups was compared using Log rank test. The multivariate analysis was performed using Cox regression model. Results: The median survival time of 131 patients was 41.6 months. The 1-year, 3-year and 5-year survival rates were 95.0%, 82.0%, and 77.0%, respectively. The 5-year recurrence-free survival rate was 81.0%, and the 5-year metastasis-free survival rate was 72.0%. Univariate analysis showed that the tumor size, initial or recurrence, surgical margin, AJCC stage, and with/without standard treatment were associated with overall survival (all P<0.05). Stratification analysis according to the American Joint Committee of Cancer (AJCC) stage showed that 5-year survival rate of stage Ⅱ patients with radiotherapy was 100.0%, which was higher than that of patients without radiotherapy (79.6%) and the difference was statistically significant (P=0.010); but no statistical significance of radiotherapy for stage Ⅲ and chemotherapy for stage Ⅱ or Ⅲ patients (all P>0.05). The multivariate analysis showed surgical margin (HR=4.220, P=0.002), with/without standard treatment (HR=4.040, P=0.030) were independent risk factors associated with prognosis of UHGPS patients. Conclusions: For UHGPS with stage Ⅱ or stage Ⅲ in extremities and trunk soft tissue, patients with complete resection and standard treatment have improved prognosis. Therefore, standard treatment, including extensive resection for the first surgery, should be performed according to expert consensus in order to increase the long-term survival rate. Adjuvant radiotherapy should be performed for stage Ⅱ patients.

[Analysis of clinicopathological characteristics and prognostic factors of foot and ankle soft tissue and bone tumors].

Objective: To evaluate the clinicopathological characteristics of foot and ankle soft tissue and bone tumor, and to analyze the prognosis and the related factors of malignant tumors in this site. Methods: 74 patients with soft tissue and bone tumors of foot and ankle from January 2006 to February 2017 were retrospectively analyzed. The clinicopathological characteristics, the treatment and survival status of malignant tumors were followed up, and the clinical and therapeutic factors related to prognosis were analyzed. Results: Of the 74 patients, 34 were males and 40 were females. The male to female ratio was 1∶1.18; the age ranged from 12 to 64 years and the median age was 42 years. Tumors located in forefoot of 22 cases, 22 in midfoot, 10 in hind foot, 14 in ankle joint and 6 in multiple sites. 14 cases were bone tumors, including 7 benign and 7 malignant, and 60 cases were soft tissue tumors, including 14 benign and 46 malignant. The most common malignant soft tissue tumors were synovial sarcomas (13 cases), and the most common benign soft tissue tumors were hemangiomas (4 cases). 44 cases of malignant tumors underwent surgery were followed up, of which were 7 bone and 37 soft tissue malignant tumors. Limb salvage surgeries were performed in 33 cases and amputation in 11 cases. The median follow-up time was 69.8 months, and the median survival time was 40.7 months. The 1-year, 3-year and 5-year survival rate of soft tissue malignant tumors was 88.0%, 73.0%, and 63.0%, respectively. The 1-year, 3-year and 5-year survival rate of bone malignant tumors was 86.0%, 57.0% and 57.0%, respectively. Univariate analysis showed that the prognostic factors affecting 5-year survival rate were tumor size and adjuvant therapy (P<0.05). Patient's gender, age, tumor location, histological type and surgical procedure had no effect on overall survival(P>0.05). Multivariate analysis showed that tumor size was an independent prognostic factor (RR=7.262, P=0.005). Conclusions: Forefoot and midfoot are more common in foot and ankle soft tissue and bone tumors. Synovial sarcoma is the most common diagnosis in malignant soft tissue tumors, and hemangioma is the most common diagnosis in benign soft tissue tumors. The prognostic factor of malignant soft tissue and bone tumors in foot and ankle is tumor size. Patients with the tumor size of 5 cm or more have a worse prognosis.

[The role of preoperative (18)F-FDG PET-CT in lymphatic metastasis diagnosis of cutaneous malignant melanoma on extremities and trunk].

Objective: To evaluate the clinical value of preoperative (18)F-Fludeoxyglucose ((18)F-FDG PET-CT) in lymphatic metastasis diagnosis of cutaneous melanoma on extremities and trunk. Methods: 112 patients with cutaneous melanoma pathologically of extremities and trunk from January 2006 to December 2016, who received (18)F-FDG PET-CT examination preoperatively, were retrospectively reviewed. The correlations between the maximal diameters of lymph nodes, the maximal standard uptake value (SUV) and the diagnostic impression grades of PET-CT examination, and the final pathological diagnosis were analyzed. The correlations between Breslow thickness of primary lesions and the diagnostic impression of PET-CT examination were also analyzed. All the above were analyzed with Receiver Operating Characteristic (ROC) curve to get the cut-off value. Based on the final results of pathological diagnosis of lymph nodes as the golden standard, the statistically significant indicators of ROC curve analysis were used to evaluate the diagnostic effect, as well as to calculate the sensitivity, specificity and accuracy. With gender, age, maximal diameter of lymph nodes, maximal SUV, diagnosis impressions, and Breslow thickness as the independent variables and pathological diagnosis results of lymph nodes as the dependent variable, two-class stepwise Logistic regression analysis was used to determine the independence of diagnostic indicators. ROC curve analysis and log rank test were used to analyze the relationship between Breslow thickness and patient survival. Results: To evaluate melanoma patients' lymph node status, the results of ROC curve analysis showed that the area under the curve of lymph node maximal diameter, maximal SUV, diagnosis impression of PET-CT examinations were 0.789, 0.786 and 0.816, respectively (all P<0.05). The cut-off values were 0.85 cm, 1.45 and 2.5, respectively. The sensitivity of the cut-off values to determine the status of lymph nodes in melanoma patients were 71.4%, 64.9% and 72.1% respectively, and the specificities were 85.2%, 88.7% and 87.0% respectively. Multivariate Logistic regression analysis showed that PET-CT diagnosis impressions had independent diagnostic significance for the lymph node status of melanoma patients (OR=11.296, 95%CI: 2.550~50.033). The area under the curve of Breslow thickness evaluating PET-CT diagnostic impression is 0.664 (P=0.042) and the cut-off value was 4.25 mm. The survival rate of the patients with Breslow thickness ≥ 4.25 mm was lower than that in the group <4.25 mm (P=0.006). Conclusions: (18)F-FDG PET-CT can help to evaluate metastases and make treatment decisions for cutaneous melanoma of extremities and trunk, especially for patients whose primary lesion's Breslow thickness has reached more than 4.25 mm. For the patients whose maximal SUV of regional lymph node is higher than 1.45 and short diameter of the largest lymph node is larger than 0.85cm, the possibility of metastases should be considered.

[Clinicopathological features and prognosis of malignant peripheral nerve sheath tumor: a retrospective study of 140 cases].

Objective: To investigate the clinicopathological features and prognosis of malignant peripheral nerve sheath tumors (MPNST). Methods: We retrospectively reviewed the clinical data of MPNST patients who were treated at Cancer Institute & Hospital, Chinese Academy of Medical Science from January 1999 to January 2016. A total of 140 patients with 66 male and 74 female with MPNST were enrolled in the study. The median age was 40 at the time of diagnosis. Survival analysis were estimated by Kaplan-Meier method and Log rank test. Multivariate analysis were estimated by Cox proportional hazards regression model. Results: The median follow-up time was 43.0 months. The 3- and 5-year overall survival (OS) rates were 56.4% and 48.6%, respectively. The 3-year local recurrence (LR) rate and distant metastasis (DM) rates were 42.9% and 49.3%, respectively. Univariate analysis showed that the tumor location, AJCC stage, S-100, radiotherapy and margin status affected 5-year OS rate (all P<0.05). The tumor location, AJCC stage, S-100, Ki-67 staining, margin status, radiotherapy and chemotherapy affected 3-year LR rate (all P<0.05). The tumor location, AJCC stage, S-100, Ki-67 staining and margin status affected 3-year DM rate (all P<0.05). Multivariate analysis showed that the tumor location, AJCC stage, S-100 were independent factors for 5-year OS rate (all P<0.05). The tumor location, Ki-67 staining and chemotherapy were independent factors for LR (all P<0.05) while the AJCC stage, margin status and Ki-67 staining were independent factors for DM (all P<0.05). Conclusions: MPSNT is an aggressive tumor with poor prognosis. Multiple factors were identified in this study. Patients with the tumor located at head and neck, advanced AJCC stage and negative S-100 usually have a low 5-year overall survival rate. Patients with the tumor located at head and neck, Ki-67 staining ≥ 20% and without chemotherapy had a higher tendency of local recurrence. Poor prognosis factors for DM were advanced AJCC stage, positive margin and Ki-67 staining ≥ 20%.

Dysregulation of Bmi1 promotes malignant transformation of hepatic progenitor cells.

Adult hepatic progenitor cells (HPCs) are involved in a wide range of human liver diseases, including hepatocellular carcinoma (HCC). Bmi1 has been reported to have vital roles in stem cell self-renewal and carcinogenesis. We have previously demonstrated that Bmi1 is upregulated in HCC with bile duct tumor thrombi, a subtype of HCC characterized by profuse expression of hepatic stem cell markers. However, the function of Bmi1 in HPCs has not yet been well elucidated. The current study was designed to investigate the effects of Bmi1 on the biological properties of rat HPCs. To accomplish this, Bmi1 was silenced or enhanced in two HPC cell lines (WB-F344 and OC3) by, respectively, using either small interfering RNA against Bmi1 or a forced Bmi1 expression retroviral vector. The biological functions of Bmi1 in HPCs were investigated through cell proliferation assays, colony-formation assays, cell cycle analysis and invasion assays, as well as through xenograft-formation assays. In this study, genetic depletion of Bmi1 repressed cell proliferation, colony formation and invasion in both assessed HPC cell lines relative to controls. Conversely, forced expression of Bmi1 in two HPCs cell lines promoted cell proliferation, colony formation and invasion in vitro. Aldehyde dehydrogenase (ALDH) assay revealed a significant increase in the number of ALDH-positive cells following the forced expression of Bmi1 in HPCs. Most importantly, transplantation of forced Bmi1 expression HPCs into nude mice resulted in the formation of tumors with histological features of poorly differentiated HCC. Taken together, our findings indicate that forced expression of Bmi1 promotes the malignant transformation of HPCs, suggesting Bmi1 might be a potential molecular target for the treatment of HCC.

Effect of p53 gene polymorphism on functions of prostate cancer cells.

Prostate cancer cells were transfected with plasmids [empty plasmids, wild-type pcDNA3.1-p53 (V/V), mutant type pcDNA3.1- p53 (G/G)] to analyze the effect of p53 gene polymorphisms on the proliferation, cycle, and apoptosis of prostatic cancer cells. Empty plasmids containing wild-type pcDNA3.1-p53 (V/V) and mutant type pcDNA3.1- p53 (G/G) were used to transfect PC3 and LNCaP cells, respectively. Cell proliferation was detected at 0, 24, 48, and 72 h using the MTT method. Cells were collected at 24 and 72 h. The distribution of cell cycles in various groups was detected using flow cytometry (propidium iodide staining method) and the apoptosis rate was detected using annexin V + propidium iodide double staining. Compared with the control group, wild-type pcDNA3.1-p53 (V/V) and mutant type pcDNA3.1-p53 (G/G) showed a significant inhibitory effect on cell proliferation (P < 0.05); the inhibitory effect of the mutant type was stronger than that of the wild-type. There was no significant difference between PC3 cells and LNCaP cells. After transfection with wild-type pcDNA3.1-p53 (V/V) and mutant type pcDNA3.1-p53 (G/G), PC3 and LNCaP cells were arrested in the G0/G1 stage. Transfection with pcDNA3.1-p53 (G/G) showed a more significant effect than transfection with pcDNA3.1-p53 (V/V). Both the wild-type pcDNA3.1-p53 (V/V) and mutant-type pcDNA3.1-p53 (G/G) led to an increased apoptosis rate of PC3 and LNCaP cells. The p53 gene polymorphism affects the proliferation, apoptosis, and cycle of prostate cancer cells and may serve as a reliable index for the diagnosis and treatment of prostate cancer.

Amiloride, a urokinase-type plasminogen activator receptor (uTPA) inhibitor, reduces proteinurea in podocytes.

This study examined the mechanism of action of amiloride, a urokinase-type plasminogen activator receptor inhibitor, in lowering proteinuria. Podocytes were resuscitated to allow for their proliferation and were observed for morphological changes. In the in vitro experiment, control, lipopolysaccharide, and lipopolysaccharide + amiloride groups were established. The expression of urokinase-type plasminogen activator receptor (uPAR) in podocytes was detected with a flow cytometer and cell motility was detected with the transwell migration assay. In the in vivo test, the urine protein volume of the model was detected at 24 h using Coomassie brilliant blue staining and the morphological changes of the podocytes were detected with immunofluorescence. The protein expression rate of uPAR in the lipopolysaccharide group was significantly higher than those in the control and lipopolysaccharide + amiloride groups (P < 0.05). The viability of cells in the lipopolysaccharide group was significantly higher than those in the control and lipopolysaccharide + amiloride groups (P < 0.05). Compared with the urine protein level in the control group at 24 h, the level in the lipopolysaccharide group increased significantly (P < 0.05), whereas compared with the urine protein level in the lipopolysaccharide group, the level in the lipopolysaccharide + amiloride group decreased (P < 0.05). uPAR expression was significantly downregulated, and the fusion of the podocyte-specific skelemin synaptopodin on the glomerulus podocytes was significantly decreased in the lipopolysaccharide + amiloride group. These results suggest that amiloride is able to reduce cell motility and thus lower proteinuria by inhibiting the expression of uPAR in podocytes.

Inactivation of Rab23 inhibits the invasion and motility of pancreatic duct adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is charac-terized by a poor prognosis and high mortality rate. In this study, we investigated the expression of Rab23 in non-tumor pancreatic tissues and PDACs via immunohistochemistry. Rab23 was found in 39 of 58 (67.2%) and in 11 of 30 (36.7%) of the PDAC and non-tumor pan-creatic tissue samples (P = 0.0073), respectively. There were signifi-cant correlations between Rab23 expression and unfavorable variables, including cancer differentiation level (P = 0.0089), lymph nodal (P = 0.0099), and distant metastases (P = 0.0173). Inactivation with small interfering RNA against Rab23 in the human pancreatic cancer cell line Panc-1 inhibited the migration and invasive potential of the cells. Our data provide new insight into the essential role of Rab23 in PDAC inva-sion and metastasis and suggest that Rab23 expression is a useful indi-cator of metastatic potential; hence, it may be a new therapeutic target for this common malignancy.

Field testing Chinese and Japanese gypsy moth nucleopolyhedrovirus and disparvirus against a Chinese population of Lymantria dispar asiatica in Huhhot, Inner Mongolia, People's Republic of China.

The activity of three geographic isolates of the gypsy moth nucleopolyhedrovirus (LdMNPV) was evaluated in field trials against larvae of the Chinese population of Lymantria dispar asiatica Vnukovskij in Inner Mongolia, People's Republic of China, in 2004, 2005, and 2006. Although the Chinese isolate of the virus, LdMNPV-H, was the most pathogenic of the isolates tested, having the lowest mean lethal concentration causing 50% and 95% larval mortality, the increase in efficacy that would be obtained by incorporating this isolate into a commercial product does not justify the time or expense required to register it for use in the United States or Canada. The commercially available North American isolate, LdMNPV-D, was moderately pathogenic, whereas the Japanese isolate, LdMNPV-J, was the least pathogenic. The slopes of the dose-response regression lines for the three virus isolates indicated that the Chinese gypsy moth larvae were more homogenously susceptible to LdMNPV-H and LdMNPV-D than to LdMNPV-J. Time-response data showed that LdMNPV-J was significantly more virulent, but at a much higher dose, than the other two isolates, causing 50% mortality in the shortest time, followed by LdMNPV-H and LdMNPV-D. Rainfall immediately after the application of LdMNPV-D in 2005 resulted in significantly reduced gypsy moth larval mortality.

Differential expression of prostaglandin E receptor subtype EP2 in rat uterus during early pregnancy.

PGE2 is essential for mammalian female reproduction. This study was to examine the expression of EP2 gene in the rat uterus during early pregnancy, delayed implantation and artificial decidualization by in situ hybridization and immunohistochemistry. There was no detectable EP2 mRNA expression in the uterus from days 1 to 4 of pregnancy (day 1 = day of vaginal sperm). A low level of EP2 immunostaining was observed in the luminal and glandular epithelium from days 1 to 4 of pregnancy. Both EP2 mRNA and protein expression were highly detected in the luminal epithelium at implantation sites on day 6 of pregnancy. EP2 expression decreased from day 7 of pregnancy and was undetectable on days 8 and 9 of pregnancy. After delayed implantation was terminated by estrogen treatment and the embryo implanted, both EP2 mRNA and protein expression were strongly observed in the luminal epithelium at the implantation site. There was no detectable EP2 expression in both control and decidualized uteri. In conclusion, these data suggest that EP2 expression at implantation site may play an important role during embryo implantation in rats.

Differential expression of peroxisome proliferator-activated receptor delta at implantation sites and in decidual cells of rat uterus.

The aim of this study was to examine the expression and regulation of peroxisome proliferator-activated receptor delta (PPARdelta) gene in rat uterus during early pregnancy by in situ hybridization and immunohistochemistry. PPARdelta mRNA expression in the luminal epithelium was high on day 1 of pregnancy, gradually declined from day 2 and was undetectable on day 5 of pregnancy. However, expression in the glandular epithelium began to increase from day 2 and was high on day 5 of pregnancy. There was no detectable PPARdelta immunostaining in the luminal and glandular epithelium from day 1 to day 5. On day 6 of pregnancy when embryos implanted, PPARdelta mRNA and immunostaining were intense in the subluminal stroma at implantation sites. On days 7 and 8, there was strong expression of both PPARdelta mRNA and intense immunostaining in the decidualized area near the lumen. There was low expression of PPARdelta in the subluminal stroma and glandular epithelium under delayed implantation. After delayed implantation was terminated by oestrogen treatment and embryo implantation was initiated, both PPARdelta mRNA and immunostaining were strongly induced in the subluminal stroma. Intense PPARdelta immunostaining was observed in the decidua under artificial decidualization, while no detectable immunostaining was seen in the uninjected control horn. Retinoid X receptor (RXRalpha) immunostaining was seen in the subluminal stroma surrounding the implanting blastocyst on day 6 and in the decidual cells on days 7 and 8 of pregnancy. In conclusion, the high PPARdelta expression at implantation sites and in the decidual cells in rat uterus indicates that PPARdelta may play an important role during implantation and decidualization.

Cloned piglets born after nuclear transplantation of embryonic blastomeres into porcine oocytes matured in vivo.

Nuclear transplantation in the pig is more difficult than in other domestic animals and only one embryonic nuclear transplantation (NT) pig has been born to date. In this study, reconstituted porcine embryos were produced by electrofusion of blastomeres from in vivo four-cell embryos to enucleated in vivo or in vitro matured (IVM) oocytes. Nuclear transfer using cumulus cells as nuclear donors was also conducted. When blastomeres were used as donors, the electrofusion rate was significantly higher in oocytes matured in vivo (91.5%) than in those matured in vitro (66.1%) (p < 0.01). After fusion, the NT embryos reconstituted from in vivo matured oocytes developed to blastocysts at a rate of 10.3% after culture in rabbit oviducts for up to 5 days, while only 5.9% of the NT embryos reconstructed from in vitro matured oocytes developed to blastocyst stage. Electrofusion rate of cumulus cell nuclei with enucleated IVM oocytes was lower (47.6%) and only 1.5% (2/136) of the reconstituted eggs developed in vitro to morula stage, and 1.9% developed to blastocysts when cultured in the ligated rabbit oviducts. Transfer of 94 embryos reconstructed by blastomere NT with in vivo matured oocytes to five synchronous recipients resulted in the birth of two cloned piglets. No piglet was born following transfer to two recipients of embryos (n = 39) derived from NT with in vitro matured oocytes. The results demonstrate that in vivo matured oocytes are better recipients than those matured in vitro for pig cloning.

Factors affecting electrofusion of 2-cell mouse embryos.

Parameters of electrofusion of 2-cell mouse embryos were optimized for application as a model for nuclear transplantation. There was considerable lysis of embryos with M(2) as the medium for fusion; however, 100% fusion (n = 58) was obtained with a single 0.31-kv / cm, 1280-musec pulse. With mannitol and sucrose solutions as the medium, a wide range of field strengths (0.31-1.41 kv / cm for 0.26 M sucrose solution and 0.31 to 2.04 kv / cm for 0.3 M mannitol solution) and durations of the electrical pulse (10-1280 musec) resulted in high rates of fusion (often 100%). Likewise, osmolarity of sucrose and mannitol solutions did not affect the rate of fusion using a 0.47-kv / cm pulse. With a field strength of 2.04 kv / cm, the proportion of embryos that fused in mannitol solution increased (P<0.05) and the proportion that were lysed decreased (P<0.05) as osmolarity increased. Both fused (162 642 , 25%) and control embryos (32 72 , 44%) continued to develop in culture for 48 h, after which they began to compact. Fused embryos were only at the 4-cell stage by this time, while control embryos were at the 8-cell stage. Optimal pulse durations are plotted for field strengths between 0.31 and 1.41 kv / cm with 0.26 M sucrose as fusion medium.