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Ceramide can be converted into sphingomyelin by sphingomyelin synthases (SMS) 1 and 2. In this study, we show that in human leukemia Jurkat cells, which express mainly SMS1, Fas ligand (FasL) treatment inhibited SMS activity in a dose- and time-dependent manner before nuclear fragmentation. The SMS inhibition elicited by FasL (1) was abrogated by benzyloxycarbonyl valyl-alanyl-aspartyl-(O-methyl)-fluoromethylketone (zVAD-fmk), a broad-spectrum caspase inhibitor; (2) did not occur in caspase-8-deficient cells and (3) was not affected in caspase-9-deficient cells. Western blot experiments showed SMS1 cleavage in a caspase-dependent manner upon FasL treatment. In a cell-free system, caspase-2, -7, -8 and -9, but not caspase-3 and -10, cleaved SMS1. In HeLa cells, SMS1 was Golgi localized and relocated throughout the cytoplasm in cells exhibiting an early apoptotic phenotype on FasL treatment. zVAD-fmk prevented FasL-induced SMS1 relocation. Thus, FasL-mediated SMS1 inhibition and relocation depend on caspase activation and likely represent proximal events in Fas signaling. FasL-induced ceramide production and cell death were enhanced in cells stably expressing an siRNA against SMS1. Conversely, in cells stably overexpressing SMS1, FasL neither increased ceramide generation nor efficiently induced cell death. Altogether, our data show that SMS1 is a novel caspase target that is functionally involved in the regulation of FasL-induced apoptosis. [ABSTRACT FROM AUTHOR]

Objective: To investigate the inplementation of cardiovascular surgery for congenital heart disease (CHD) in China. Methods: A cross-sectional study was carried out. The CHD cardiovascular surgery data collected by the Chinese Society of Extracorporeal Circulation from 2017 to 2021 in 31 provinces (autonomous regions/municipalities) of China were retrospectively reviewed, the implementation of CHD cardiovascular surgery in different provinces, regions, general/specialized hospitals, and different age groups (whether≤18 years old) were summarized, and the correlation analysis between the number of surgeries carried out in each province/region and the gross regional product and the number of the regional population was performed. Results: Between 2017 and 2021, the annual volume of CHD cardiovascular surgery was 77 120, 77 634, 81 161, 62 663 and 71 492, respectively, showing a decreasing trend. Meanwhile, the proportion of CHD patients aged≤18 years who underwent cardiovascular surgery also showed a downward trend, from 79.8% (61 557/77 120) in 2017 to 58.6% (41 871/71 492) in 2021 ( P =0.027). The number of surgical cases varied greatly among different provinces, including 4 provinces with≥5 000 cases and 9 provinces with 2 000-5 000 cases. In the five years, the number of CHD cardiovascular surgeries in Central and East China was the largest, accounting for 41.1%-45.5% of the total surgical cases. The proportion of CHD surgery cases≤18 years old was the highest in Southwest China (69.7%-87.4%) and the lowest in Northeast China (28.2%-68.9%). Except for 2021, the number of cases carried out by each region between 2017 and 2020 was correlated with the gross regional product ( r =0.929, 0.929, 0.893 and 0.964, respectively, all P <0.05) and the population ( r =0.821, 0.893, 0.821 and 0.857, respectively, all P <0.05). Hospitals that performed more than 100 operations (20.5%±1.2% of the total number of hospitals) completed 86.2%±1.2% of the total number of operations in China during the 5-year period. In 2017 and 2021, the number of CHD cardiovascular surgeries preformed in children's/women's and children's specialized hospitals accounted for 24.3% (18 772/77 120) and 23.8% (17 012/71 492) of the total number of cases in China, respectively. Conclusions: From 2017 to 2021, the number of cardiovascular surgery for CHD decreases slightly, but the proportion of surgery for adult CHD patients increases significantly.There is a strong correlation between the number of CHD operations in each region and their economic development status. The scale of CHD cardiovascular surgery performed in children's hospitals/women's and children's hospitals accounts for about a quarter of the total volume in China.

Objective: To examine the clinical value of fluorescence-guided indocyanine green (ICG) laparoscopic anatomical hepatectomy in the treatment of primary hepatocellular carcinoma. Methods: Data from patients diagnosed with hepatocellular carcinoma and who underwent laparoscopic hepatectomy with ICG fluorescence navigation in the Department of Liver Surgery and Liver Transplantation Center of West China Hospital between September 2020 and May 2022 were retrospectively collected. There were 53 males and 19 females, with an age of (55.5±12.9)years(range:42.6 to 68.4 years). Among them, 13 of the cases underwent laparoscopic anatomical liver resection(LALR) guided by tans-arterial ICG,43 of the cases received LAIR guided by portal vein negative ICG, and 16 of the cases received LALR positive by portal vein. Comparison among the three groups was performed by one-way ANOVA; and the rank sum test was used for comparison between groups. The counting data was expressed as percentage,and the χ 2 test or Fisher's exact probability method was used for comparison between groups. Results: (1) Postoperative pathology: Resection R0 was achieved in all operations. The maximum tumor diameter of the patients in the arterial staining group, the reverse staining group, and the positive staining group( M (IQR)) was 2.5 (2.4) cm, 3.0 (2.5) cm and 3.0(2.4) cm,respectively. There were no statistically significant differences in the maximum tumor diameter between the three groups ( P =0.364). The minimum tumor margin was 1.1 (1.1) cm, 1.0 (1.0) cm, 1.1 (1.6) cm in the the arterial staining group, reverse staining group and the positive staining group, respectively. There was no significant difference in the margin among the three groups ( P =0.878). (2) Operation conditions: the operation time of the arterial staining group, the negative staining group, and the positive portal staining group was (348±93)minutes,(277±112)minutes,and (295±116)minutes,respectively. There were no significant differences in operation time among the three groups ( P =0.134). The intraoperative blood loss of the three groups was 80(150)ml,200(350)ml,and 100(150)ml,respectively. There was no statistically significant difference in intraoperative bleeding volume between the three groups( P =0.743). All cases were not transfused during the operation and were not converted to laparotomy. ALT in the arterial staining group was higher than in the negative staining group in the first two days after the operation ((559±398)IU/L307(257) IU/L, q =235.5, P =0.004;(611±389)IU/L(331±242) IU/L, q =265.2, P =0.002). There was only one case of a grade III complication (Clavien-Dindo grading system) postoperative complication in the negative and positive staining group of the portal vein, respectively. Tumor markers in all patients decreased to the normal range after 2 months of operation. Conclusion: Laparoscopic anatomical hepatectomy guided by ICG fluorescence through arterial staining and portal vein staining is safe and feasible for primary hepatocellular carcinoma treatment.

Objective: The aim of the study was to explore the influencing factors of exercise tolerance in patients with myocardial infarction (MI) after percutaneous coronary intervention (PCI) revascularization in acute state. Methods: A total of 112 patients with first MI undergoing PCI revascularization in acute state and completing cardiopulmonary exercise testing (CPET) were enrolled. Exercise capacity was evaluated by peak oxygen consumption percentage (VO(2 peak)%) in CPET. Patients were divided into normal exercise capacity (NEC) group ( n= 40) and abnormal (AEC) group ( n= 72) according to VO(2 peak)% value. Clinical manifestations, histories of hypertension and diabetes, medications, coronary arterial angiography and echocardiography findings of patients were compared. The onsets of diabetes and blood glucose levels during the period of CPET were evaluated in the MI patients with diabetes. The patients were followed up for major adverse cardiovascular events (MACE) (admission due to chest pain, re-revascularization, re-infarction and all-cause death) within 24 months after PCI. Multivariate logistic regression analyses were conducted to examine influencing factors for exercise tolerance. Results: The ratio of diabetes, type C lesions in the AEC group were higher than those in the NEC group (diabetes: 37.5% vs. 17.5%; type C lesions: 69.4% vs. 42.5%, respectively, all P< 0.05). The left ventricular ejection fraction (LVEF) in patients in the AEC group was lower than that in the NEC group [(60.6±10.0)% vs. (65.0±8.2)%, P= 0.019]. Multivariate logistic regression analyses showed that history of diabetes and history of type C lesions were the independent risk factors for the declined exercise capacity in the MI patients after PCI revascularization ( OR= 3.14, 95 %CI 1.167-8.362, P= 0.023; OR= 3.32, 95 %CI 1.444-7.621, P< 0.01). Among the MI patients with diabetes, the duration of diabetes in the AEC group was significantly longer than that in the NEC group[(7.7±3.6)years vs. (5.0±2.4)years] and the proportions of subjects reaching target levels of fasting plasma glucose (40.7% vs. 57.1%) and glycosylated hemoglobin A1c(HbA1c) (55.6% vs. 71.4%) in this group were significantly lower than those in the NEC group (all P< 0.05). A multivariate logistic regression analysis showed that reaching HbA1c target was an independent predictor of improved exercise tolerance in MI patients with diabetes who received PCI ( OR= 2.518, 95 %CI 1.395-7.022, P= 0.021). No significant differences were observed in incidence of admission due to chest pain, re-revascularization and re-infarction between the two groups within 24 months after PCI between the groups. Conclusions: Diabetes and type C lesions are independent risk factors of declined exercise capacity in patients with first myocardial infarction who received revascularization in acute state. Reaching target HbA1c is independent factor of improved exercise capacity in patients with myocardial infarction and diabetes.

Objective: To investigate the association between the internal exposure levels of phthalic acid esters (PAEs) and executive function (EF) of preschool children. Methods: Between October 2008 and October 2010, pregnant women who accepted pregnancy health care services in four municipal medical and health institutions in Ma'anshan city, Anhui Province, were recruited as study objects. A total of 5 084 pregnant women and 4 669 singleton live births were enrolled in this cohort. The follow-up study was conducted from April 2014 to April 2015. A total of 3 725 data-completed preschool children aged 3 to 6 years older entered in this study. The method of analysis seven metabolites of phthalates in urine was high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and divided objects into low ( P (0)- P (24)), medium ( P (25)- P (74)) and high ( P (75)- P (100)) groups according to their exposure concentrations. To investigate the executive function of preschool children, we used the Behavior Rating Inventory of Executive Function-Preschool Version (BRIEF-P). Univariate and multivariate statistical method was used to analyze the etiology association between the phthalate metabolites levels and preschool children's executive function. Results: In this study, 53.6% (1 997/3 725) of preschoolers were boys, children's age was (51.5±5.6) months. The detection rates of seven phthalate esters were: mono-n-methyl phthalate (MMP) was 99.89% (3 721/3 725); mono-ethyl phthalate (MEP) was 99.97% (3 724/3 725); mono-benzyl phthalate (MBzP) was 69.10% (2 574/3 725); mono-butyl phthalate (MBP), mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP) and mono- (2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) were 100.00% and mono-ethylhexyl phthalate (MEHP) was 99.95% (3 723/3 725). The median concentrations of the seven phthalate metabolites were: 17.71, 15.36, 0.07, 155.24, 10.73, 14.67, 3.59 µg/L, respectively. The median concentrations corrected by urinary creatinine were 29.65, 26.65, 0.12, 257.73, 17.94, 24.80, 6.27 µg/g Cr, respectively. The P (25) and P (7)5 concentration of the total PAEs metabolites corrected by urinary creatinine were 1.20 µmol/g Cr and 3.04 µmol/g Cr. After adjusted the relevant demographic information: children sex, children age in month, maternal age, parental education levels, household exposure to secondhand smoke and whether the child is the only child as confounds, multivariate logistic regression model showed that the risk of inhibitory self-control index (ISCI) dysplasia in MEHHP high concentration group and MEOHP high concentration group were 1.71 and 1.54 times ( OR= 1.71, 95 % CI: 1.11-2.62; OR= 1.54, 95 % CI: 1.01-2.34) than in low concentration group. The risk of ISCI dysplasia in total PAEs metabolites high concentration group was 1.55 times ( OR= 1.55, 95 % CI: 1.00-2.38) than in low concentration group. Conclusion: Phthalates exposure may damage the executive function of preschool children.

Objective: To summarize the experience of totally thoracoscopic cardiac surgical (TTCS) at congenital heart diseases (CHD) treatment., Methods: From April 2000 to March 2016, 2 543 patients with CHD underwent TTCS in Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, including 957 male and 1 586 female patients. The age ranged from 0.5 to 66.0 years with a mean age of (21±18) years. The body weight ranged from 6 to 118 kg with a mean of (49±30) kg. Patients were diagnosed with echocardiography or transesophagel echocardiography as complex or complicated CHD which was unsuitable for percutaneous procedure. Surgical procedures were performed through 3 holes inserted at the right chest wall, and catheters were placed in the right femoral artery and vein to set up cardiopulmonary bypass.The ascending aorta was cross-clamped with long tailor-made forceps and the myocardium was protected by coronary perfusion with cold crystalloid(blood)cardioplegia. There were 787 cases (from January 2013 to December 2015) were selected to compare with 710 cases underwent conventional thoracotomy over the same period. Statistical analysis was performed by t test, t' test, rank-sum test, χ(2) test and Fisher exact test, respectively., Results: The total death rate and the major complication rate of the operation were 0.35% (9/2 543) and 2.28% (58/2 543), respectively. All patients were followed up 1 to 190 months and the average follow-up time was (75±34) months. No residual shunt or obvious mitral/tricuspid regurgitation was observed, and the patients gained better cardiac function as level Ⅰ to Ⅱ (New York Heart Association classification). There was no significant difference in aorta clamp time, ICU stay, hospital cost, and surgical fatality rate between 787 patients underwent TTCS and 710 conventional thoracotomy. The cardiopulmonary bypass time ((31±20) minuets vs. (40±17) minuets, t'=9.407, P=0.001), operation time ((91±27) minuets vs. ( 102±64) minuets, t'=4.251, P=0.000), hospital stay ((5.3±2.2) d vs. (13.0±4.0) d, t'=45.463, P=0.000), postoperative drainage (M(QR): 33(17) ml vs. 121(53) ml, T=2.632, P=0.000) and major complications (7/787 vs. 23/710, χ(2)=10.49, P=0.001) were significantly reduced and no sternal deformation occurrence (0 vs. 192/710, χ(2)=244.10, P=0.000) in TTCS group. While the cost was higher in TTCS group ((24 000±400) yuan vs. (20 000±400) yuan, t=19.320, P=0.000)., Conclusion: TTCS is feasible, safe, and minimal invasive for patients with CHD, resulting in quick recovery and good median-long term outcomes.

Chemokines have been shown to be expressed in some malignant or precancerous tissues. However, the role of these chemokines on tumor development or progression is not clear. The expression patterns of chemokines in gastric cancer tissues were examined in 86 surgically resected samples using immunohistochemistry. Macrophage inflammatory protein-1beta (MIP-1beta) was clearly detected in many gastric carcinoma cells. In most of the differentiated carcinomas, intracellular localization of MIP-1beta was detected in more than 5% of cancer cells, although the percentages of MIP-1beta-positive cells differed among each sample. Undifferentiated carcinomas showed contrasted staining pattern between solid type and non-solid (diffuse) type. MIP-1beta was totally absent in all the poorly differentiated carcinomas with solid type growth pattern (por1). In contrast, MIP-1beta was highly expressed in all of the non-solid type of poorly differentiated carcinoma (por2) and signet-ring cell carcinoma samples. In particular, MIP-1beta was strongly stained in carcinoma cells at the front of invasive lesions. In 43 diffuse type undifferentiated cancers, tumors with high expression of MIP-1beta exhibited significantly more lymph node metastasis. Our results suggest a possibility that MIP-1beta may be related to the scattering and invasion step of gastric carcinoma cells with undifferentiated phenotype.

The exposure of human stomach cancer KATO III cells to oolong tea polyphenol extact [OTPE] containing oolong tea polyphenol trimer [OTP trimer] as a main component, led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with OTPE. The fragmentation by OTPE of DNA to oligonucleosomalsized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These data suggest that the growth inhibition by OTPE results from the induction of apoptosis in the KATO III cells.

TCR engagement of immature CD4(+)CD8(+) thymocytes induces clonal maturation (positive selection) as well as clonal deletion (negative selection) in the thymus. However, the cell death execution events of thymocytes during the negative selection process remain obscure. Using a cell-free system, we identified two different DNase activities in the cytosol of in vivo anti-TCR-stimulated murine thymocytes: one that induced chromosomal DNA fragmentation, which was inhibited by an inhibitor of caspase-activated DNase, and another that induced plasmid DNA degradation, which was not inhibited by an inhibitor of caspase-activated DNase. We purified the protein to homogeneity that induced plasmid DNA degradation from the cytosol of anti-CD3-stimulated thymocytes and found that it is identical with cyclophilin B (Cyp B), which was reported to locate in endoplasmic reticulum. Ab against Cyp B specifically inhibited the DNA degradation activity in the cytosol of anti-CD3-stimulated thymocytes. Furthermore, recombinant Cyp B induced DNA degradation of naked nuclei, but did not induce internucleosomal DNA fragmentation. Finally, we demonstrated that TCR engagement of a murine T cell line (EL4) with anti-CD3/CD28 resulted in the release of Cyp B from the microsome fraction to the cytosol/nuclear fraction. Our data strongly suggest that both active caspase-activated DNase and Cyp B may participate in the induction of chromosomal DNA degradation during cell death execution of TCR-stimulated thymocytes.

Two new ichthyotoxic compounds, aspidin PB (8) and dryofragin (9), along with three known phloroglucinol derivatives (1-3) and five terpenoids, were isolated from the whole herbs of Dryopteris fragrans by toxicity-directed fractionation using Oryzias latipes (Japanese name; medaka). The structures of the new compounds were determined by spectroscopic methods including 2D NMR techniques. Amongst the isolates, aspidin PB (8), dryofragin (9), and 1-5 exhibited potent ichthyotoxic activity against medaka with a median tolerance limit (TLm after 24 h) of 1.2-4.3 microg/ml. These compounds which are toxic to fish also had a potent inhibitory effect on the activation of Epstein-Barr virus early-antigen (EBV-EA) induced by tetradecanoyl phorbol 13-acetate, which is an in vitro short-term assay for anti-tumor promoting agents. Aspidin BB (2) and albicanol (4), which exhibited strong inhibitory effects on the EBV-EA activation, significantly suppressed an in vivo two-stage carcinogenesis on mouse skin.

Recombination activating gene-1 (RAG-1) and RAG-2 are expressed in lymphoid cells undergoing the antigen receptor gene rearrangement. A study of the regulation of the mouse RAG-2 promoter showed that the lymphocyte-specific promoter activity is conferred 80 nucleotide (nt) upstream of RAG-2. Using an electrophoretic mobility shift assay, it was shown that a B-cell-specific transcription protein, Pax-5, and a T-cell-specific transcription protein, GATA-3, bind to the -80 to -17 nt region in B cells and T cells, respectively. Mutation of the RAG-2 promoter for Pax-5- and GATA-3-binding sites results in the reduction of promoter activity in B cells and T cells. These results indicate that distinct DNA binding proteins, Pax-5 and GATA-3, may regulate the murine RAG-2 promoter in B and T lineage cells, respectively. (Blood. 2000;95:3845-3852)

The epidemiological survey of prevalence of NIDDM (non-insulin dependent diabetes mellitus) and IGT(impaired glucose tolerance) was conducted among 9450 residents aged 25-70 in some areas of Hubei Province, China. The results show that NIDDM and IGT prevalences are 2.62% and 4.48%, respectively. There is no significant difference between male and female (P > 0.05). The NIDDM prevalence in cities is slightly higher than that in countryside, but the difference is not significant (P > 0.05). However, the IGT prevalence in city is significantly higher than that in countryside (P < 0.01). The prevalence of both NIDDM and IGT is increasing along with the age of the population. It is also significantly related to the family history of NIDDM, hypertension, and high body mass index (BMI). By using stepwise logistic regression to analyse the risk factors of NIDDM, age (OR = 1.86), BMI(OR = 2.69), family history (OR = 2.84) and hypertension (OR = 2.23) entered the model (significance level is alpha = 0.05).

We isolated and characterized several monoclonal antibodies against a protein complex containing the flagellar movement-initiating phosphoprotein (MIPP) that appears to play a crucial role in the initiation of flagellar movement in quiescent spermatozoa of Salmonid fish. The effects of the antibodies on the phosphorylation of MIPP, as well as on the initiation of movement, in model sperm cells were studied. Three monoclonal antibodies, namely, FMI7, FMI18, and FMI27, were found specifically to inhibit both the initiation of flagellar movement and the phosphorylation of MIPP. These antibodies did not recognize denatured MIPP; they only recognized the native antigen. FMI7 exclusively recognized the denatured form of a 38-kDa protein, which may possibly be a protein kinase responsible for the phosphorylation of MIPP. Immunofluorescence analysis in situ of model sperm cells with the antibodies showed that the antigen was localized predominantly in the basal structure of the spermatozoon. Thus, the results clearly demonstrate the involvement of MIPP in the initiation of flagellar movement and the control of flagellar motility.

Phosphorylation of a tyrosine residue within a 15-kDa protein has been found to play a role in the initiation of flagellar movement of quiescent spermatozoa of the rainbow trout, Salmo gairdneri [Morisawa & Hayashi (1986) Biomed. Res. 6, 181-184; Hayashi et al. (1987) J. Biol. Chem. 262, 16692-16698]. In order to find a more accessible source of the 15-kDa protein for biochemical studies, phosphorylation of proteins was studied in other organs from other species. Cell-free extracts from spermatozoa and testes of Salmonidae were prepared and all catalyzed the cAMP-dependent, Mg(2+)-requiring, but Ca(2+)-independent phosphorylation of the 15-kDa proteins. Protein from the testes of a white salmon, Oncorhynchus keta, was isolated by conventional purification methods. The 15-kDa protein in the cell-free extract was found to be complexed with several other proteins such that the 15-kDa protein and its phosphorylating activity were copurified. Isolation of a sufficient amount of the complex for the preparation of antibodies against the various constituents is now possible.

Nuclear DNA content of 131 pancreatic duct epithelial lesions, including 10 normal ducts, 30 intraductal proliferations with mild atypia (groups I-II), 30 with moderate atypia (group III), 24 with severe atypia (group IV), 14 of carcinoma in situ (group V), and 23 invasive carcinomas, was analyzed using microspectrophotometry. DNA histograms were classified into diploid, polyploid and aneuploid patterns. All of normal duct epithelia showed diploidy. Polyploid patterns were observed in 3 (10%) lesions of groups I-II, 17 (56.7%) of group III, 14 (58.4%) of group IV, 7 (50%) of group V, and 6 (26.1%) of invasive carcinomas, and aneuploid patterns were observed in 0%, 10%, 33.3%, 50% and 73.9%, respectively. This distribution of ploidy patterns revealed a gradual shift to the main ploidy from diploid to polyploid followed by aneuploid in proportion to the increase of the degree of epithelial atypia. The frequencies of polyploid cells in each lesion were determined. Their averages were 0.2% in groups I-II, 1.9% in group III, 3.4% in group IV, 4.4% in group V, and 6.7% in invasive carcinoma. The S+G2M phase fractions were significantly higher in proliferative epithelia than in normal. The results of this study suggest that duct epithelial proliferations of the pancreas have "genetic instability" leading to a serial clonal evolution and play a significant role in the progression of pancreatic duct cell carcinoma.