[Can tumor vaccines really cure liver cancer?]

In China, the incidence of liver cancer remains high. Approximately 80% of diagnosed patients are in the intermediate and advanced stages, with a high recurrence rate and poor prognosis after surgery. Therefore, substantially reducing the incidence and mortality has always been a major clinical challenge for liver cancer. In recent years, immune checkpoint inhibitor therapy represented by programmed death protein 1 (PD-1) antibody is gradually innovating the traditional paradigm of tumor treatment, but the beneficiary population in liver cancer patients is relatively limited. With the rapid development of high-throughput sequencing, proteomics and immunomics and other fields, the demand for precision medicine continues to increase. Tumor vaccines, especially derived from neoantigens, have shown promising therapeutic effects in malignant solid tumors such as melanoma and non-small cell lung cancer due to their immunogenicity. Combining the latest research reports at home and abroad, this paper emphasis on whether tumor vaccines can effectively treat or even cure liver cancer.

[Clinicopathological features of myeloid sarcoma and DLBCL in the breast: a comparative study].

Objective: To study the clinicopathological features, diagnosis and differential diagnosis of myeloid sarcoma of the breast. Methods: Ten cases of myeloid sarcoma (MS) and 19 cases of diffuse large B cell lymphoma (DLBCL) of the breast were selected from Peking University People's Hospital from February 2005 to September 2019. The cases were evaluated by microscopy and immunohistochemistry basing on WHO classification (2008 and 2017). Results: For the 10 cases of MS, the mean and median age was 33.8 and 31 years (range 23 to 47 years) respectively. All patients presented with breast masses; six presented with B symptoms (6/10); and LDH level was elevated in four patients. The largest tumor dimension was 1.0 to 5.3 cm (mean 2.7 cm). All 10 patients had history of acute myeloid leukemia (AML), and in one patient, the AML occurred after chemotherapy for hydatidiform mole. One case was classified as M0, four were M2, two were M4 and three were M5. For the AML, all patients received chemotherapy and nine were treated by allogeneic hematopoietic stem cell transplant (allo-HSCT) and the breast masses occurred4 months to 2 years post-transplant. Using Ann Arbor staging, five cases were stage Ⅰ, three were stage Ⅱ, and 2 were stage Ⅳ. The MS was found in the left breast (two cases); right breast (three cases) and both breasts (five cases). Lymphocyte in peripheral blood, B symptom and site of lesion had statistical significance between myeloid sarcoma and DLBCL(P<0.05). The tumor cells were primitive, expressing MPO, CD43, CD117, etc. All ten patients had follow-up information, and the median survival period was 14.4 months (range 1 to 50 months). Seven patients died. The prognosis of patients with MS was worse than DLBCL(P=0.002). Conclusions: The clinical history, pathologic morphology, immunophenotyping and molecular studies are very important for diagnosing MS tumors in the breast, and MS may occur after allo-HSCT for AML. Tumor resection, chemotherapy, radiotherapy and donor lymphocyte infusion are recommended for treatment. The prognosis is poor.

[Clinical characteristics of patients with delayed hepatolenticular degeneration].

Objective: To evaluate the clinical manifestations, metal metabolism, imaging characteristics and treatment response in patients with delayed Wilson disease (WD). Methods: Patients with untreated WD (40 with delayed onset and 40 with non-delayed onset) were enrolled. Twenty healthy people were included as normal controls. All patients were evaluated with modified Young scale neural symptom scores, grade of Child liver function and mental symptoms rating scale, magnetic resonance imaging (MRI) scan, magnetic sensitive imaging (susceptibility weighted imaging, SWI), metal metabolism. Corrected phase (CP) was measured at SWI. After 2 week treatment, neurologic symptoms, liver function, and metal metabolism were reviewed. Results: The total score of neurological symptoms in WD patients with delayed onset was lower than that of non-delayed onset (13.00±6.87 vs. 21.13±5.53, P=0.033). The scores of SCL-90 and HAMA depression scales in patients with delayed onset were lower than those of non-delayed onset. On T(2) weighted imaging, areas including substantia nigra and thalamus, the caudate nucleus, globus pallidus, putamen presented high signal rate in patients with delated onset than those with non-delayed (P=0.022, 0.037, 0.022, 0.037, 0.029 respectively). The SWI CP values of cangbai sphere and shell nucleus in patients with delayed onset were lower than those with non-delayed onset. Patients with delayed onset had higher urinary copper than those with non-delayed onset before and after treatment (P=0.040, 0.036). After treatment, the score of abnormal tremor and gait in patients with delayed onset was decreased (P=0.037, 0.044), while as the occurrence of neurological symptoms was increased by 10%, and the liver function level in patients with delayed WD was decreased in 3 cases. Conclusions: The brain of WD patients with delayed onset is mainly composed of metal deposits, however the cell damage is not apparent. Clinical symptoms are characterized by significant liver injury, but relatively mild neurological and psychiatric symptoms. Patients with delayed WD have higher urinary copper excretion than those with non-delayed WD. Chelating agents improves the neurological symptoms in patients with delayed onset.

[Analysis of proliferative lesions of haematopoietic and lymphoid tissue in the female productive tract].

Objective: To study the clinicopathologic features, diagnosis and differential diagnosis of tumors of haematopoietic and lymphoid tissue in the female productive tract. Methods: Eleven cases of myeloid sarcoma and leukemia, 9 of non Hodgkin lymphoma (NHL) , 13 of cervical lymphoma-like lesions were selected from Peking University People's Hospital from January 2006 to August 2017. According to WHO classification of tumors of haematopoietic and lymphoid tissues (2008) and updated classification(2016), the cases were studied by microscopy, immunohistochemistry and in situ hybridization. Results: In 20 cases of tumors of haematopoietic and lymphoid tissue, the mean and median age was 48.5 and 56 years old (range: 16-77 years old) . In cases of lymphoma-like lesion of uterine cervix, the mean and median age was 45.9 and 48 years old (range: 23-62 years old) . The patients with neoplasm present as fever, fatigue, hypogastralgia, colporrhagia and mass etc. Eight cases had history of acute myeloid leukemia, and 3 had myeloid leukemia while pregnancy. One case of chronic lymphocytic leukaemia/small lymphocytic lymphoma (CLL/SLL) had history of ovary small cell carcinoma and high grade serous carcinoma resected with chemotherapy. One case of diffuse large B cell lymphoma (DLBCL) had history of renal transplantation. Lactic dehydrogenase (LDH) was elevated in 9 cases (9/18) . The cases of lymphoma-like lesion present as contact bleeding in most cases and all located in cervix. Four cases of neoplasm located in vulva, 1 in vagina, 4 in cervix, 4 in uterine corpus, 8 in ovary and 2 in placenta. Clinical staging of NHL: 4 case was stageⅠ, 1 case of stageⅢ, and 4 cases of stage Ⅳ. Pathological morphology: 9 cases were myeloid sarcoma, 2 cases were placenta invaded by myeloid leukemia. Six cases were DLBCL, and 1 case was CLL/SLL, 1 case was mucosa associated lymphoid tissuse lymphoma (MALToma) , and 1 case was anaplastic large cell lymphoma. Resected mass, chemotherapy was performed in tumors of haematopoietic and lymphoid tissue. Five cases of myeloid sarcoma and 2 of NHL died. In 13 cases of lymphoma-like lesion of uterine cervix, the general condition was good as following up. Conclusions: The clinical history, pathological morphology and immunohistochemistry are very important for diagnosing tumors of haematopoietic and lymphoid tissue in the female productive tract. Resection with chemotherapy is recommended in treatment. The prognosis of lymphoma-like lesion of uterine cervix is good, and should be differentiated from lymphoma.

Coexistence of Eph receptor B1 and ephrin B2 in port-wine stain endothelial progenitor cells contributes to clinicopathological vasculature dilatation.

BACKGROUND: Port-wine stain (PWS) is a vascular malformation characterized by progressive dilatation of postcapillary venules, but the molecular pathogenesis remains obscure. OBJECTIVES: To illustrate that PWS endothelial cells (ECs) present a unique molecular phenotype that leads to pathoanatomical PWS vasculatures. METHODS: Immunohistochemistry and transmission electron microscopy were used to characterize the ultrastructure and molecular phenotypes of PWS blood vessels. Primary culture of human dermal microvascular endothelial cells and in vitro tube formation assay were used for confirmative functional studies. RESULTS: Multiple clinicopathological features of PWS blood vessels during the development and progression of the disease were shown. There were no normal arterioles and venules observed phenotypically and morphologically in PWS skin; arterioles and venules both showed differentiation impairments, resulting in a reduction of arteriole-like vasculatures and defects in capillary loop formation in PWS lesions. PWS ECs showed stemness properties with expression of endothelial progenitor cell markers CD133 and CD166 in non-nodular lesions. They also expressed dual venous/arterial identities, Eph receptor B1 (EphB1) and ephrin B2 (EfnB2). Co-expression of EphB1 and EfnB2 in normal human dermal microvascular ECs led to the formation of PWS-like vasculatures in vitro, for example larger-diameter and thick-walled capillaries. CONCLUSIONS: PWS ECs are differentiation-impaired, late-stage endothelial progenitor cells with a specific phenotype of CD133+ /CD166+ /EphB1+ /EfnB2+ , which form immature venule-like pathoanatomical vasculatures. The disruption of normal EC-EC interactions by coexistence of EphB1 and EfnB2 contributes to progressive dilatation of PWS vasculatures.

[Tumors of lymphoid and hematopoietic tissue of spleen: a clinicopathologic analysis of 53 cases].

Objective: To study the clinicopathologic features, diagnosis and differential diagnosis of the tumors of lymphoidand hematopoietic tissue of the spleen(TLTS). Methods: Fifty-three cases of TLTS were selected from the pathologic files from Peking University People's Hospital from April 2002 to April 2017. According to WHO classification of tumors of hematopoietic and lymphoid tissues (2008) and its updated classification (2016), the cases were studied by microscopy, immunohistochemistry and in situ hybridization, combined with the bone marrow biopsy and clinical examination. Results: In 53 cases of TLTS, the male to female ratio was 3.4∶1.0; the mean age was 55.4 years (range 21-76 years), and all patients presented with variable degree of splenomegaly. Laboratory examination showed increased percentage of lymphocyte in peripheral blood in 22 cases, and elevated serum LDH level in 24 cases. Abnormal blood counts were seen in 26 cases pre-operatively, in which 22 cases showed complete or partial correction of these abnormalities post-operatively (84.6%, 22/26). The clinical symptoms included abdominal pain or distension, fatigue, fever, and weight loss, etc. Seventeen cases presented with lymphadenopathy of abdomen or other sites. Bone marrow biopsy was performed in 30 cases, and 19 cases were involved by tumor (63.3%). Of all 53 cases, 43 were diagnosed as primary splenic lymphoma (PSL), and the remaining 10 cases as secondary TLTS. According to Ann Arbor staging, 14 cases were stages Ⅰ or Ⅱ, 6 were stage Ⅲ and 28 were stage Ⅳ. By histopathologic classification, 43 cases of PSL were splenic B-cell marginal zone lymphoma (SMZL; 48.8%, 21/43), diffuse large B cell lymphoma (DLBCL; 23.3%, 10/43), splenic diffuse red pulp small B-cell lymphoma (11.6%, 5/43), mantle cell lymphoma (9.3%, 4/43), follicular lymphoma (4.7%, 2/43), and composite lymphoma (CL, DLBCL and classical Hodgkin lymphoma; 2.3%, 1/43). The remaining 10 cases were chronic lymphocytic leukaemia/small lymphocytic lymphoma (4 cases), hairy cell leukaemia (1 case), hepatosplenic T-cell lymphoma (HSTL; 5 cases), with lesions in other sites. Of the 53 cases of TLTS, 47 were B cell neoplasm (88.7%, 47/53), and the T cell neoplasms were all HSTL(5 cases, 9.4%, 5/53), 1 case was composite lymphoma. In 11 cases of TLTS, EBER in situ hybridization was performed and all cases were negative. Forty eight cases had follow-up data, and the median survival period was 17.0 months(range: 1-96 months). The survival of patients with SMZL and DLBCL were 25.7 and 18.6 months respectively. Thirteen patients died (27.1%, 13/48). The prognosis of those with elevated LDH level, high clinical stage, B symptoms and older than 60 years of age was worse. And the prognosis of DLBCL was worse than that of SMZL. There was no statistically significant difference between these factors and prognosis (P>0.05). Conclusions: Most TLTS cases present with splenomegaly and abnormal blood counts, and complete or partial remission of blood counts isseen after splenectomy. The most common pathologic types of TLTS are SMZL and DLBCL. Definite diagnosis of TLTS could be made by combining clinical features, histopathology, immunophenotype, genetics, bone marrow biopsy and laboratory examination.

Quantitative trait loci mapping of the stigma exertion rate and spikelet number per panicle in rice (Oryza sativa L.).

The stigma exertion rate is a polygenic inherited trait that is important for increased seed yield in hybrid rice breeding. To identify quantitative trait loci (QTL) associated with high stigma exertion rate, we conducted QTL mapping using 134 recombinant inbred lines derived from XieqingzaoB and Zhonghui9308, which have high and low stigma exertion rates, respectively. A total of eight QTLs (qSES6, qSSE11, qDSE1a, qDSE1b, qDSE10, qDSE11, qTSE1, and qTSE11) for single stigma exertion, double stigma exertion, and total stigma exertion were detected. The locations of qSSE11 and qTSE11 have not been previously reported, and the qDSE11 allele from parent XQZB exhibited a positive additive effect. In addition, three QTLs (qSNP1, qSNP3a, and qSNP3b), for spikelet number per panicle were identified. Of note, one QTL (qSNP1) was detected in two different environments (Hainan and Zhejiang). To evaluate the advantage of exerted stigma for cross-pollination, single, dual, and total stigma exertion should be considered separately for future genetic improvement in the production of rice hybrid seeds. In addition, this study provides information for fine mapping, gene cloning, and marker assisted selection, with emphasis on the latter.

Comparison of gene regulatory networks of benign and malignant breast cancer samples with normal samples.

The aim of this study was to explain the pathogenesis and deterioration process of breast cancer. Breast cancer expression profile data GSE27567 was downloaded from the Gene Expression Omnibus (GEO) database, and breast cancer-related genes were extracted from databases, including Cancer-Resource and Online Mendelian Inheritance In Man (OMIM). Next, h17 transcription factor data were obtained from the University of California, Santa Cruz. Database for Annotation, Visualization, and Integrated Discovery (DAVID)-enrichment analysis was applied and gene-regulatory networks were constructed by double-two-way t-tests in 3 states, including normal, benign, and malignant. Furthermore, network topological properties were compared between 2 states, and breast cancer-related bub genes were ranked according to their different degrees between each of the two states. A total of 2380 breast cancer-related genes and 215 transcription factors were screened by exploring databases; the genes were mainly enriched in their functions, such as cell apoptosis and proliferation, and pathways, such as p53 signaling and apoptosis, which were related with carcinogenesis. In addition, gene-regulatory networks in the 3 conditions were constructed. By comparing their network topological properties, we found that there is a larger transition of differences between malignant and benign breast cancer. Moreover, 8 hub genes (YBX1, ZFP36, YY1, XRCC5, XRCC4, ZFHX3, ZMAT3, and XPC) were identified in the top 10 genes ranked by different degrees. Through comparative analysis of gene-regulation networks, we identified the link between related genes and the pathogenesis of breast cancer. However, further experiments are needed to confirm our results.

[Clinical characteristics and prognostic risk factors analysis of carbapenem-resistant organism in the department of hematology].

Objective: To study the distribution and drug resistance of Carbapenem-Resistant Organism (CRO) and to analysis the risk factors of CRO 30-day mortality. Methods: A total of 181 patients with CRO infection diagnosed in Department of Hematology, Fujian Medical University Union Hospital from January 2018 to June 2020 were retrospectively investigated. The clinical and laboratory data of the patients were collected, the prognosis of patients diagnosed with CRO infection in day 30 was followed up, and the risk factors of prognosis were analyzed. The clinical significance of Carbapenem-Resistant Enterobacteriaceae (CRE) active screening was further evaluated in the CRE subgroup. Results: Among the total of 181 CRO isolates, 47.2% were CRE, 37.0% were Pseudomonas aeruginosa, and 32.6% were Klebsiella pneumoniae, which were highly resistant to carbapenem and had high MIC value, 76.8% (139/181) of CRO were MIC of imipenem resistance≥16 µg/ml. The main sources of isolates were blood and sputum. The 30-day all-cause mortality rates of patients with CRO or CRE infection were (41.4±3.7) % and (44.7±5.4) %, respectively. The COX multivariate regression analysis showed that the level of procalcitonin >0.2 ng/ml and the MIC value of imipenem resistance ≥ 16 µg/ml were independent risk factors for 30-day mortality of CRO infected patients. The CRE subgroup analysis showed that MIC value of imipenem resistance ≥16 µg/ml were independent risk factors for 30-day mortality of CRE infected patients. The 30-day cumulative survival rate of patients with CRE active screening was higher than the patients without CRE active screening [ (68.0±9.3) % vs (50.0±6.5) %, P=0.21]. Conclusion: The high MIC value of imipenem resistance isolates seriously affects the prognosis of patients with CRO infection in the hematology department, and the mortality rate was high. CRE active screening is expected for early prevention, early diagnosis, and early treatment for high-risk patients.

Homocysteine induced oxidative stress in human umbilical vein endothelial cells via regulating methylation of SORBS1.

OBJECTIVE: The aim of the present study was to investigate the mechanism of homocysteine (Hcy) induced oxidative stress in the human umbilical vein endothelial cells (HUVECs). PATIENTS AND METHODS: The HUVECs were isolated from umbilical vein vascular wall of 12 patients and treated with Hcy. The malondialdehyde (MDA) level was measured using the thiobarbituric acid (TBA) method. The expressions of superoxide dismutase 2 (SOD2), endothelial nitric oxide synthase (eNOS), and intercellular adhesion molecule 1 (ICAM-1) were detected by Western blot and RT-PCR. The genome-wide DNA methylation assay was performed using the Infinium Human Methylation 450 BeadChip. The specific DNA methylation was determined using bisulfite sequencing analysis. To evaluate the role of sorbin and SH3 domain-containing protein 1 (SORBS1), the HUVECs were transfected with small interfere RNA (siRNA) targeting SORBS1 (SORBS1-siRNA). RESULTS: Hcy induced MDA level in HUVECs, and increased ICAM-1 expression both in protein and mRNA levels. The protein and mRNA levels of SOD2 and eNOS were inhibited by Hcy induction. However, the effects of Hcy on MDA level and expressions of SOD2, eNOS, and ICAM-1 were attenuated by folic acid (Fc) and vitamin B12 (B12) treatment. DNA total methylation level in Hcy treated cells was significantly decreased compared to the control group, while the DNA total methylation levels were increased after treatment with Fc and B12. The methylation level of SORBS1 in Hcy treatment group was higher than that of control group. And the methylation level of SORBS1 induced by Hcy was attenuated by Fc and B12 treatment. SORBS1-siRNA transfection induced the MDA levels and reduced the expressions of SOD2 in HUVECs. CONCLUSIONS: We indicated that Hcy induced oxidative stress in HUVECs via regulating methylation of SORBS1. We also found that Fc and B12 treatment attenuated the oxidative stress and inflammation induced by Hcy in HUVECs. The findings indicated that Fc and B12 might be effective agents for the treatment of Hcy induced AS.

A Comparison of Plain Radiography with Computer Tomography in Determining Coronal and Sagittal Alignments following Total Knee Arthroplasty.

Introduction: Optimal coronal and sagittal component positioning is important in achieving a successful outcome following total knee arthroplasty (TKA). Modalities to determine post-operative alignment include plain radiography and computer tomography (CT) imaging. This study aims to determine the accuracy and reliability of plain radiographs in measuring coronal and sagittal alignment following TKA. Materials and Methods: A prospective, consecutive study of 58 patients undergoing TKA was performed comparing alignment data from plain radiographs and CT imaging. Hip-knee-angle (HKA), sagittal femoral angle (SFA) and sagittal tibial angle (STA) measurements were taken by two observers from plain radiographs and compared with CT alignment. Intra- and inter-observer correlation was calculated for each measurement. Results: Intra-observer correlation was excellent for HKA (r>0.89) with a mean difference of <1.9°. The least intra-observer correlation was for SFA (mean r=0.58) with a mean difference of 8°. Inter-observer correlation was better for HKA (r>0.95) and STA (r>0.8) compared to SFA (r=0.5). When comparing modalities (radiographs vs CT), HKA estimations for both observers showed the least maximum and mean differences while SFA observations were the least accurate. Conclusion: Radiographic estimation of HKA showed excellent intra- and inter-observer correlation and corresponds well with CT imaging. However, radiographic estimation of sagittal plane alignment was less reliably measured and correlated less with CT imaging. Plain radiography was found to be inferior to CT for estimation of biplanar prosthetic alignment following TKA.

Posterior condylar cartilage thickness and posterior condylar offset of the femur: a magnetic resonance imaging study.

PURPOSE: To measure the femoral posterior condylar cartilage (PCC) thickness and the posterior condylar offset (PCO) and determine the correlation between the 2 parameters in 530 normal subjects using magnetic resonance imaging (MRI). METHODS: Records of 287 male and 243 female patients (mean age, 40.7 years) who underwent MRI for traumatic soft tissue knee injuries not involving the femoral PCC and did not have symptomatic knee arthritis were reviewed. RESULTS: The PCC thickness was comparable in the lateral and medial sides (2.04 vs. 1.99 mm, p=0.13). Males had thicker PCC in the medial (2.05 vs. 1.92 mm, p=0.0006) and lateral (2.16 vs. 1.86, p<0.0001) sides than females. Age did not correlate with PCC thickness. The bony PCO was larger in the medial than lateral side (25.8 vs. 22.6 mm, p<0.0001). Males had a larger PCO than females in the medial side only (26.1 vs. 25.5 mm, p = 0.0195). The bony PCO did not correlate with PCC thickness. CONCLUSION: Femoral PCC thickness was comparable in the medial and lateral sides. Males had thicker PCC in the medial and lateral sides than females.

Clinical and financial benefits of intra-articular tranexamic acid in total knee arthroplasty.

PURPOSE: To determine whether intra-articular tranexamic acid (TXA) use after total knee arthroplasty (TKA) results in decreased postoperative blood transfusion and length of hospital stay. METHODS: Medical records of 1981 patients (mean age, 69.2 years) who underwent primary TKA with (n=1006) or without (n=975) TXA use by any of 4 knee arthroplasty surgeons were reviewed. TXA (3000 mg/30ml) was administered via an epidural catheter into the knee joint after wound closure. Postoperative blood transfusion was given to patients with haemoglobin (Hb) level <80 g/dl on days 1 and 2 or with symptoms of acute anaemia. RESULTS: Intra-articular TXA use after TKA resulted in a lower blood transfusion rate (4.5% [45/1006] vs. 14.8% [144/975], p<0.0001), fewer units of blood transfused (86 vs. 313 units, p<0.0001), fewer units of blood transfused per 100 patients (8.5 vs. 32.1, p<0.0001), and shorter length of hospital stay (4.7±2.3 vs. 5.3±2.7 days, p<0.0001). Total cost savings with respect to the reduction in blood transfusion was AU$143.68 per patient. When the change in length of hospital stay and TXA costs were included, the overall saving was AU$631.36 per patient. CONCLUSION: Intra-articular TXA use can reduce costs as a result of decreased blood transfusion rate and length of hospital stay in patients undergoing TKA.

Prostaglandin F2alpha stimulates the Raf/MEK1/mitogen-activated protein kinase signaling cascade in bovine luteal cells.

Upon binding to its G protein-coupled transmembrane receptors, the actions of PGF2alpha on the corpus luteum are initiated by the phospholipase C/diacylglycerol-inositol 1,4,5-trisphosphate (InsP3)/Ca2+-protein kinase C (PKC) pathway. However, little is known about the downstream intracellular signaling events that can lead to transcriptional activation in response to PGF2alpha. The present study was conducted to examine the involvement of the mitogen-activated protein kinase (MAPK) signaling cascade in the corpus luteum. Three isoforms of the Raf family of oncoprotein kinases (A-Raf, B-Raf, and Raf-1 or c-Raf) were detected in bovine luteal cells. Raf-1 and B-Raf, but not A-Raf, were activated by PGF2alpha (1 microM) and the pharmacological PKC activator phorbol myristate acetate (PMA, 20 nM). Kinetic analysis revealed that PGF2alpha rapidly and transiently activated Raf-1. In vitro protein kinase assays demonstrated that activation of Raf-1 and B-Raf resulted in the phosphorylation and activation of MAPK kinase (MEK1), which subsequently phosphorylated p42mapk. As determined by hyperphosphorylation, tyrosine phosphorylation, and enzymatic activity, p42mapk and p44mapk were rapidly and transiently activated by both PGF2alpha (1 microM) and PMA (20 nM). Additionally, both PGF2alpha (1 microM) and PMA (20 nM) stimulated phosphorylation of Raf-1, MEK1, and p42mapk in 32P-labeled cells. Our data demonstrate that PGF2alpha activates the Raf/MEK1/p42/44mapk signaling cascade in bovine luteal cells and that the actions of PGF2alpha are mimicked by the PKC activator PMA. Activation of the Raf/MEK1/MAPK signaling cascade by PGF2alpha in luteal cells provides a mechanism to transduce signals initiated by PGF2alpha receptors on the cell surface into the nucleus. Activation of the Raf/MEK1/MAPK signaling cascade may be associated with transcriptional activation of luteal genes possessing activator protein-1-binding sites.

An investigation of pulsatile release tablets with ethylcellulose and Eudragit L as film coating materials and cross-linked polyvinylpyrrolidone in the core tablets.

To develop new pulsatile release tablets, which can suppress drug release in stomach and release the drug rapidly after a predetermined lag time of about 3 h in intestine, the use of tablets with ethylcellulose/Eudragit L as a coating film and cross-linked polyvinylpyrrolidone in the core tablets was investigated. The release of diltiazem hydrochloride (DIL) as a model drug in the core tablets was investigated in vitro. The lag time (t10) was prolonged with an increase of the coating level, whereas the drug release rate was almost constant, irrespective of the coating level. The water-uptake study and electron microscope photographs suggested the mechanism of pulsatile release of drug. Pulsatile release tablets containing 60 mg DIL with 4.4 h of lag time (t10) in vitro were administrated to eight volunteers. The mean plasma concentration curves showed 4.9 h of lag time (tlag), 8.0 h of time to maximum concentration (tmax) and 3.1 h of time between tmax and tlag (t(psi)) in vivo. Relative bioavailability was 1.05 for pulsatile release tablets compared to conventional tablets.

Insulin-like growth factor I retards apoptotic signaling induced by ethanol in cardiomyocytes.

Apoptosis of cardiac muscle cells may contribute to the development of cardiomyopathy and heart failure. Alcohol (ethanol) abuse is a major cause of cardiomyopathy, but its underlying mechanism remains unknown. To determine whether ethanol causes apoptosis in cardiac muscle and whether insulin-like growth factor I (IGF 1) improves cardiac muscle survival upon ethanol exposure, we have defined the effects of ethanol and IGF I in primary cardiomyocytes. Ethanol decreased cell viability in dose-response manner from 0.2% to 1%. In contrast, ethanol (0.2-1%) did not alter viability of cardiac fibroblasts. To assess the occurrence of apoptosis, DNA fragmentation was determined with quantitation of nucleosomes. Nucleosomes were increased in ethanol-treated cells, thus confirming the apoptotic effects of ethanol. The pro-apoptotic Bax protein and Caspase 3 are important proteins of apoptotic signaling. The content of Bax and the activities of Caspase 3 were increased upon ethanol exposure. IGF I partially suppressed Bax induction, Caspase 3 activation, DNA fragmentation, and increased cardiomyocyte survival. The effects of IGF I on ethanol-induced apoptosis can be inhibited with a chemical inhibitor of PI 3 Kinase (LY-294002), suggesting that anti-apoptotic actions of IGF I involves PI 3 Kinase. These results may have important implications on further understanding the pathogenesis of alcoholic heart disease and the development of new strategies to treat alcoholic cardiomyopathy.

Bioconcentration, elimination and metabolism of 2,4-dinitrotoluene in carps(Cyprinus Carpio L.).

Bioconcentration curves of 2,4-dinitrotoluene(2,4-DNT) in carps (whole fish, liver, intestine and muscle) were investigated using semistatic system. For whole fish, its curve could be described as a gentle peak which began with a rise in concentration to summit or steady state, then declined and reached lower level followed by another steady state. For liver and intestine, their curves both contained two successive peaks, with the second peak followed by slight fluctuation. Bioconcentration factors of 2,4-DNT in whole fish during the first and second steady state were 9.15 and 4.15,(97.86 and 44.39, based on lipid content), respectively. By logarithmic plotting, two straight-lines with different slopes(3.6 and 0.1 d-1) were measured for elimination. According to peaky curves of 2,4-DNT in whole fish, liver and intestine, smaller BCFs than calculated BCFs based on the regression equations for inert chemicals, and large rate constants of elimination, biotransformation was inferred to have happened in tissues such as liver, intestine, and other tissues. Two metabolites were separated from liver and identified as 4-amino-2-nitrotoluene(4A2NT) and 2,4-diamino-toluene(2,4-DAT) on HPLC, their retention times were 23.1 and 8.8 min, respectively. In bioconcentration test of 2,4-DNT in liver, two metabolites and parent were determined at the same time at intervals, higher concentrations of 4A2NT and 2,4-DAT were found when level of 2,4-DNT declined. Such results demonstrated our inference that metabolism caused the declines in bioconcentration curves. A one-compartment model was set up to simulate the bioconcentration, in which biotransformation adhered to Delayed Enzyme-Catalytic Logarithmic Kinetics. Good fit of model curves with measured values could be observed.

Flexor digitorum profundus tendon avulsion through a recurrent enchondroma--a case report.

Flexor digitorum profundus (FDP) tendon avulsion from the distal phalanx is a well recognised injury, which usually follows a hyperextension force to a flexed distal interphalangeal (DIP) joint. It is commonly seen in contact sport athletes, with a predilection for the ring finger.(2,4) Avulsion of the FDP tendon from pathological bone is an infrequent occurrence. It has, however, been reported to occur in association with an enchondroma of the distal phalanx.(2,3) To our knowledge, an FDP tendon avulsion through a recurrent enchondroma has not been reported. We present the case findings of such an event.