[Analysis of prognosis factors of postoperative cardiac complications in colorectal cancer patients with comorbid coronary artery disease].

Objective: To examine the prognosis factors of postoperative cardiac complications in colorectal cancer patients co-morbidated with coronary artery disease. Methods: Clinical data of 449 patients colorectal cancer patients co-morbidated with coronary artery disease accepted redical surgery from April 2013 to April 2020 at Department of General Surgery, Peking Union Medical College Hospital were analyzed retrospectively. There were 306 males and 143 females, aging (68.7±8.9) years (range: 44 to 89 years). Postoperative acute coronary syndrome, new-onset arrhythmia and heart failure that causes clinical symptoms were recorded as cardiac complications. t test, χ2 test and Fisher exact test were used for univariate analysis of prognosis factors of postoperative cardiac events. The variables with P<0.05 were included in the multivariate Logistic regression was used to determine the independent prognosis factors. Results: After surgery, 44 patients (9.8%) suffered from at least one cardiac event, including 30 patients with acute coronary syndrome, 19 patients with new-onset arrhythmia and 9 patients with heart failure. There were 3 deaths in the cohort within 30 days after surgery. Two patients died from cardiac-related complications, and one from septic shock due to postoperative anastomotic leaks. On Univariate analysis showed that cardiac complications were associated with age ≥80 years, co-morbidated diabetes, emergency surgery, re-operation, anastomotic leakage, intestinal flora disorder and elevation of preoperative neutrophil-lymphocyte ratio (χ2: 4.308 to 12.219, all P<0.05). Multivariate Logistic regression analysis identified age ≥80 years(OR=3.195, 95%CI: 1.379 to 7.407, P=0.007), co-morbidated diabetes (OR=2.551, 95%CI: 1.294 to 5.025, P=0.007), emergency surgery (OR=4.717, 95%CI: 1.052 to 20.833, P=0.043), and elevated preoperative neutrophil-lymphocyte ratio (OR=1.114, 95%CI: 1.018 to 1.218, P=0.018) as independent prognosis factors for cardiac complications. Conclusions: Emergency surgery, advanced age, co-morbidated type 2 diabetes and elevated preoperative neutrophil-lymphocyte ratio may increase the risk of postoperative cardiac complications in colorectal cancer patients with coronary artery disease. Surgeons should strictly master surgical indications, pay attention to preoperative assessment, perioperative monitoring, and diagnosis and treatment of postoperative complications in order to reduce the risk of complications.

[A nationwide survey of acute pancreatitis:a report from 126 hospitals in China].

Objective: To investigate the current status of acute pancreatitis(AP) diagnosis and treatment in hospitals of different levels in China. Methods: A cross-sectional survey was conducted. The Acute Pancreatitis Diagnosis and Treatment Practice Questionnaire was designed and sent to the members of the Group of Pancreatic Surgery Chinese Society of Chinese Medical Association Branch and some other hospitals online from 8th to 24th December, 2020. Observation indicators included general information, AP diagnosis and assessment, treatment strategies, follow-up information, and comparisons of clinical practice between 3A-level and non-3A-level hospitals were performed. Counting data was used χ 2 test or Fisher exact test. Results: A total of 126 valid questionnaires were collected in final analysis, of which 75.4% (95/126) were from 3A-level hospitals, 15.9%(20/126) and 8.7%(11/126)were from other third-level and second-level hospitals,respectively. Of all participants, 88.1% (111/126)used classic AP diagnostic criteria, and 88.1% (111/126)conducted severity assessment. The revised Atlanta classification and determinant-based classification were commonly used, accounting for 72.1%(80/111) and 22.5%(25/111), respectively. 70.6%(89/126)used predictive models, including APACHE Ⅱ score, imaging models(modified CT severity index or Balthazar scoring) and Ranson criteria. For patients with early pancreatic or peripancreatic infection, 75.4%(95/126) preferred antibiotic therapy, and for those with infected walled-off necrosis, 61.1% (77/126) preferred percutaneous catheter drainage.When surgical intervention required,preferred methods were laparoscopic transabdominal surgery(37.3%, 47/126) and open surgery(25.4%,32/126). 61.1%(77/126) accepted "delayed surgery" notion. 32.5%(41/126) routinely used the step-up approach. For mild biliary acute pancreatitis, 44.4%(56/126) underwent cholecystectomy during the same hospital admission. Regarding follow-up, ideal overall follow-up periods were 6 months(46.0%,57/124) and 12 months(33.1%, 41/124), and follow-up interval was 3 months(50.8%,63/124) and 1 month(23.4%, 29/124). Comparing clinical practice of AP between 3A-level hospitals and non-3A-level hospitals, we found that the former had a significantly higher proportion of annual AP admission number of over 100(34.7%(33/95) vs.12.9%(4/31), χ 2=5.372, P=0.020), and higher proportion of routine severity assessment(68.4%(65/95) vs. 35.5%(11/31), χ²=11.107, P=0.004), higher proportion of routine severity prediction(45.3%(43/95) vs. 12.9%(4/31), χ²=13.549, P=0.001). When surgical intervention required, the proportion of step-up approach was significantly higher(37.9%(36/95) vs.16.1%(5/31), χ 2=8.512, P=0.017). Significantly more participants preferred that follow-up should be completed by full-time staff(35.8%(34/95) vs. 22.6%(7/31), χ²=8.154, P=0.043) in 3-A level hospitals. Conclusions: The standardization of AP diagnosis is relatively high in China. However, standardized assessment of severity and prediction need to be further prompted, especially in non-3A-level hospitals. Regarding AP treatment, especially the minimally invasive intervention strategy would be the focus of the promotion of standardized AP practice in the future.

Neonatal sevoflurane anesthesia induces long-term memory impairment and decreases hippocampal PSD-95 expression without neuronal loss.

AIM: Volatile anesthetics are widely used in the clinic, and sevoflurane is the most prevalent volatile anesthetic in pediatric anesthesia. Recent findings question the potential risks of volatile anesthetics on brain development. Evidence suggests that sevoflurane may cause neuronal deficiency. This study investigates the long-term effect of sevoflurane in the developing brain. MATERIALS AND METHODS: We anesthetized 7 day-old rats for 4 h with 2.5% sevoflurane. A Morris water maze was used to evaluate hippocampal function 7 weeks after sevoflurane exposure. Nissl staining was performed to analyze neuronal loss. PSD-95 (postsynaptic density protein-95) expression in the hippocampus was measured using a western blot. RESULTS: The exposure to 2.5% sevoflurane caused long-term deficits in hippocampal function and decreased hippocampal PSD-95 expression without neuronal loss. This study demonstrates that P7 rats exposed for 4 h to 2.5% sevoflurane have significant spatial learning and memory impairment 7 weeks after anesthesia. In addition, PSD-95 expression in the hippocampus decreased at P56 without neuronal loss. CONCLUSIONS: These data suggest that sevoflurane causes neurotoxicity in the developing brain, which may be attributed to decreased PSD-95 in the hippocampus.

Synemin isoforms in astroglial and neuronal cells from human central nervous system.

The intermediate filament (IF) synemin gene encodes three IF proteins (H 180, M 150, L 41 kDa) with overlapping distributions. Synemin M was present early with vimentin and nestin. Synemin H was found later in the nervous system and mesodermic derivatives concomitantly with angiogenesis and the migration of neural crest cells. Synemin L appeared later in neurons. A series of in vitro cell cultures were done to identify the linkage between synemin isoforms and specific cell types of the central nervous system (CNS). The neurons and glia from the brains of humans and rats were cultured and double immunostaining done with antibodies against the H/M or L synemin isoforms and neural cell types (betaIII-tubulin or NeuN) or astrocyte intermediate filaments (GFAP or vimentin). In neurons of the CNS, synemin H/M were co-expressed with GFAP, vimentin or nestin in glial cells, whereas synemin L was found in neurons.

Different expression of synemin isoforms in glia and neurons during nervous system development.

The synemin gene encodes proteins belonging to the intermediate filament family. These proteins confer resistance to mechanical stress and modulate cell shape. Three synemin isoforms, of 180 (H), 150 (M) and 41 (L) kDa, are produced by alternative splicing of the pre-mRNA and are regulated differently during development. The three isoforms differ in their C-terminal tail domains, while their IF rod domains are identical. Synemins H/M occurred together with nestin and vimentin in glial progenitors during the early differentiation of the developing mouse central nervous system. They are later found in GFAP-labeled cells. In contrast, the L isoform appeared only in neurons, together with neurofilaments and betaIII-tubulin in the brain after birth. However, synemin L appeared from E13 in the peripheral nervous system, where it was confined to the neurons of spinal ganglia. In the meantime, the synemin H/M isoforms were found in both the neurons and Schwann cells of the sensorial ganglia from E11. Tissue fractionation and purification of IFs from adult mouse spinal cord revealed that the synemin L isoform binds to neurofilaments associated with the membrane compartment. This report describes the synthesis of the three synemin isoforms by selective cell types, and their temporal and spatial distributions. Mechanisms specific to neurons and glia probably control the splicing of the common synemin mRNA and the synthesis of each synemin isoform.

The mouse synemin gene encodes three intermediate filament proteins generated by alternative exon usage and different open reading frames.

We have previously cloned and characterized the human synemin gene, which encodes two intermediate filament proteins (IFPs). We now show that the mouse synemin gene encodes three different synemin isoforms through an alternative splicing mechanism. Two of them, synemin H and M are similar to human alpha and beta synemin, and the third isoform, L synemin, constitutes a new form of IFP. It has a typical rod domain and a short tail (49 residues) with a novel sequence that is produced by a different open reading frame. The synthesis of H/M synemins starts in the embryo, whereas the synemin L isoform is present in adult muscles. The H/M isoforms are bound to desmin or vimentin in the muscle cells of wild-type mice. Using desmin- and vimentin-deficient mice, we have obtained direct evidence that synemin is associated with muscle intermediate filaments in vivo. The organization of the synemin fibril is disrupted in skeletal and cardiac muscle when desmin is absent and in smooth muscle when vimentin is absent. The fact that the three synemin isoforms differ in the sequences of their tail domains as well as in their developmental patterns suggests that they fulfill different functions.

The expression of Quox 1, a homeodomain-containing protein, in sympathetic ganglion cells is regulated in vitro by growth factors.

Quox 1, a quail homeobox gene, is the first vertebrate Antp-type homeobox gene to be described that is expressed in the forebrain. We have already shown that the Quox 1 protein is specifically expressed in post-mitotic sensory neurons. A subpopulation of sympathetic ganglion cells was also found to be labelled by anti-Quox 1 in vitro, but it is not clear whether this protein is expressed in sympathetic ganglion cells in vivo and, if so, the conditions which regulate its expression in vitro. In the present study, we used immunocytochemistry to find out whether Quox 1 expression in sympathetic ganglion cells in vitro is regulated by environmental signals. We found that several peptide growth factors can regulate Quox 1 expression in cultured sympathetic ganglion cells, and that they do so at physiological concentration and in a variety of ways. Basic fibroblast growth factor (FGF-2) induces Quox 1 protein expression, whereas insulin and human insulin-like growth factor-I (IGF-I) down-regulate Quox 1 expression.

[Screening of mouse connexin 31 genomic clone and mapping by fluorescence in situ hybridization].

Connexin 31(Cx31) is one of the human genetic deafness disease genes. To determine the structure and function of Cx31 gene of mouse, the mouse Cx31 gene clone was screened from the genomic library with the probe obtained according to the sequence of the mouse Cx31 gene cDNA. Cx31 gene was mapped to the center of mouse chromosome 4 by fluorescence in situ hybridization. The present study lays a foundation for researches on structure and function of Cx31 gene, as well as establishment of the relative transgenic mice and gene knockout mice.

Effect of human neuronal tau on denaturation and reactivation of rabbit muscle D-glyceraldehyde-3-phosphate dehydrogenase.

Human neuronal tau-40 (htau-40) has been used to study denaturation and renaturation of rabbit muscle D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12). Inactivation of GAPDH incubated with tau was more distinguishably detected than that of control GAPDH during thermal and guanidine hydrochloride (GdnHCl) denaturation. However, tau did not influence the activity of GAPDH at room temperature or in solution without GdnHCl. A marked change in both the emission intensity and emission maximum of the intrinsic fluorescence at 335 nm of GAPDH with tau was observed when GdnHCl concentration was 0.8 M, but that of the control without tau occurred in 1.2 M GdnHCl. The first-order rate of the decrease in the fluorescence intensity of the enzyme with tau was approximately twice as great as that of GAPDH without tau. Kinetics of inactivation of GAPDH with tau in 0.2 M GdnHCl was a monophasic procedure, instead of the biphasic procedure followed by the control, as described before [He, Zhao, Yan and Li (1993) Biochim. Biophys. Acta 1163, 315-320]. Similar results were obtained when the enzyme was thermally denatured at 45 degrees C. It revealed that tau bound to the denatured GAPDH but not the native molecule. On the other hand, tau suppressed refolding and reactivation of GAPDH when this enzyme was reactivated by dilution of GdnHCl solution. Furthermore, tau improved the aggregation of the non-native GAPDH in solutions. It suggested that tau acted in an anti-chaperone-like manner towards GAPDH in vitro. However, tau lost that function when it was aggregated or phosphorylated by neuronal cdc2-like protein kinase. It showed that tau's anti-chaperone-like function depended on its native conformation.

Isolation of quail qMEF2D gene and its expression pattern in the developing central nervous system.

We report here the identification of the first avian MEF2 gene, termed qMEF2D. qMEF2D is the first MEF2 protein that contains 41 repeats of glutamine in the C-terminal. This quail gene is more abundantly expressed, in a transient fashion, in the developing brain than in the muscle cells.

[The expression of Quox-1 gene homologous sequence in the development of early human embryos].

By using the b2 fragment of Quox-1 gene as probe, we have confirmed that the Quox-1 gene homologous sequence exists in the human genome according to the results of Southern blot. Studies on the expression of Quox-1 homologous sequence in early human embryos from 26 to 37 days by means of immunohistochemistry technigue with Quox-1 protein antibodies showed the spatiotemporal expression patterns: in 26 days embryo Quox-1 homologous sequence was expressed in many places including neural tube, but 30 days later, tits expression sites were limited to notochord, digestive epithelium, myotome, cardiac muscle cell and periderm. The functions in control and regulation of Quox-1 gene homologous sequence during the early development of human embryo were discussed.

Spatial and temporal effects of axial structures on myogenesis of developing somites.

To elucidate the precise roles of axial structures in the myogenic differentiation of the somite, we have examined the effects of the axial organs' precise spatial position during migration and differentiation of somitic cells by using in vivo transplantation of the neural tube and of the notochord directly into the paraxial mesoderm. Differentiation of myotomal cells was identified through the use of Quox 1 antibody which recognizes specifically a quail homeoprotein Quox 1. We have demonstrated that both ectopic neural tube and notochord are able to influence the myogenesis in somites, but that the spatial position of axial organs and the degree of somite maturation at grafting time are decisive. At the level of the somites which were already formed and developmentally advanced (somites III-VI), both neural tube and notochord promote myogenesis, and the promoting effect of notochord is more efficient than that of the neural tube. In the newly formed somites (I-II) and/or the segmental plate mesoderm, the notochord inhibits the myogenesis of somites, whereas the neural tube plays an evident myogenic promoting role. But the myogenic effect of the neural tube depends not only upon the stage of developing somites and presomitic mesoderm, but also on the developmental maturation of the neural tube. We have demonstrated that the myogenic effect of the rostral part of neural tube is stronger than that of its caudal part. This observation suggests that there is a gradient of myogenic effect along the rostrocaudal axis of the neural tube, which depends on the developmental maturation of neural tube, and that the generation of skeletal muscle during somitogenesis may be in relation with the rostrocaudal gradient of the capacity of the neural tube to stimulate myogenesis since somites are also distributed along an anteroposterior axis.

Quox-1, an Antp-like homeobox gene of the avian embryo: a developmental study using a Quox-1-specific antiserum.

Quox-1 is an homeogene isolated in the quail by using the Drosophila Antp cDNA as a probe. Quox-1 homeodomain sequence homology is 98% and 100% with Antp and mouse Hox1.1 homeodomains respectively. The region of the molecule 3' to the homeodomain has no homology with any known protein sequence (Xue et al., 1991). We have raised an antiserum, designated anti-Quox-1, to the C-terminal portion of the quail Quox-1 protein. By Western blot analysis the polyclonal antibody detects proteins of the same apparent molecular weight in quail and chick embryo extracts. The developmental expression of the Quox-1 protein was examined by immunocytochemistry in whole-mount preparations or on sections of quail and chick embryos and in cell cultures. The same binding patterns were observed in the two species examined. The initial expression of Quox-1 protein was detected in the neural primordium at presomitic stages. The immunoreactivity is concentrated in the head fold and progressively extends over the fore- and midbrain when the neural tube closes. Later in development, the Quox-1 protein becomes widespread over the whole central nervous system, including forebrain and eyes. The Quox-1 gene is also active in a subpopulation of neural crest cells at the early phase of their migration in vivo and in vitro. Later on, the sensory neurons of the DRG showed anti-Quox-1 immunoreactivity, whereas sympathetic ganglia did not express this gene. Expression of Quox-1 in somitic mesenchyme is at first detected in virtually all cell nuclei of the early segmented somites, but after sclerotome-dermomyotome dichotomy, Quox-1 expression becomes restricted to the myotome. These results show that in contrast to the other vertebrate class I Hox genes, whose homeobox is of Antp type, Quox-1 has a domain of expression that includes fore- and midbrain, as well as the rest of the neural anlage.

Expression of the gene encoding tyrosine hydroxylase in a subpopulation of quail dorsal root ganglion cells cultured in the presence of insulin or chick embryo extract.

Avian sensory ganglia contain a population of normally latent autonomic precursors with catecholaminergic potentialities. The present study examines the expression of the tyrosine hydroxylase (TH) gene in quail dorsal root ganglia (DRG) by both in situ hybridization and polymerase chain reaction (PCR) techniques. In situ hybridization using quail TH cDNA as a probe demonstrated the presence in DRG cell cultures of TH mRNA in a subpopulation of cells that never express the adrenergic phenotype in vivo. Expression of the TH gene in autonomic precursor cells of DRG in culture is totally dependent on the presence either of insulin or chick embryo extract. The numbers of catecholaminergic cells expressing TH mRNA and TH immunoreactivity evolve in a closely similar manner during the culture period. Using two primers, specific for highly conserved 5' regions of TH cDNA, it was possible to detect the same band of DNA amplified by PCR in total RNA from DRG cultures grown in the presence of insulin, sympathetic ganglia and adrenal gland. No amplified DNA was detected in uncultured DRG cells. These data further indicate that, under the influence either of insulin or a still unknown factor contained in the CEE, the TH gene is induced in a subpopulation of DRG cells.

Quox-1, a quail homeobox gene expressed in the embryonic central nervous system, including the forebrain.

This paper reports the cloning and sequencing of a quail homeobox-containing gene, Quox-1, and its expression pattern in embryos from 3 to 6 days (E3 to E6) of development as determined by in situ hybridization. The opening reading frame in cDNA clone g11 corresponds to a predicted protein of 242 amino acids. Quox-1 protein displays high sequence similarity to the Antennapedia family, especially to the mouse homeodomain-containing protein Hox-1.1 (100% identity in the homeobox region, 77% at the 5' end beyond the homeobox). However, the carboxyl-terminal domain of the postulated protein has no significant homology with other known homeoproteins, including Hox-1.1. In situ hybridization experiments showed that Quox-1 is widely expressed in the developing central nervous system including the entire brain and the spinal cord. Outside the central nervous system, transcription of Quox-1 was mainly detected in the endoderm-derived epithelium of esophagus, trachea, and other digestive organs, as well as in the sensory epithelium of the olfactory region and perichondrium of the vertebrae. Thus, Quox-1 transcripts have a remarkably wide distribution that, unlike the other vertebrate homeobox genes examined to date, encompasses the rostral part of the developing nervous system, including the forebrain.

Insulin and insulin-like growth factor-I can trigger the differentiation of catecholaminergic precursors in cultures of dorsal root ganglia.

Avian sensory ganglia contain a population of normally latent autonomic-type precursors with noradrenergic potentialities. Their differentiation in vitro into cells expressing tyrosine hydroxylase immunoreactivity is acutely dependent on the presence of one or more substances found in chick embryo extract (CEE). We have used cultures of dissociated dorsal root ganglia from embryonic quail as a model system in which to assay factors promoting catecholaminergic differentiation, the latter being appreciated quantitatively in terms of the number of tyrosine hydroxylase-positive cells present after 6 days in vitro; over a large range of concentrations, the number of such cells is directly proportional to the amount of CEE in the medium. In the course of attempts to replace CEE by defined bioactive molecules, we found that epidermal growth factor, fibroblast growth factor or nerve growth factor possessed negligible, or only marginal, noradrenergic differentiation-promoting activity. In contrast, insulin, at nanomolar levels, triggered expression of the catecholaminergic phenotype as well as did CEE. Insulin-like growth factor-I, at similar concentrations, had an analogous effect. It is suggested that an insulin-like molecule may play a role in the normal differentiation of sympathoblast precursors in vivo.

High-affinity uptake of noradrenaline in quail dorsal root ganglion cells that express tyrosine hydroxylase immunoreactivity in vitro.

During embryonic life, avian sensory ganglia contain cells with the potential to express, under appropriate experimental conditions, a number of properties characteristic of autonomic sympathetic neurons. Thus, cells capable of synthesizing noradrenaline (NA) from tyrosine differentiate when dorsal root ganglia (DRG) from 10-15 d embryonic quail are grown in culture (Xue et al., 1985a, b). In the present study, we show that cultures of DRG from 10 d embryos can take up 3H-NA by a high-affinity (Km = 1.0 microM), temperature-dependent process that can be inhibited by desmethylimipramine. By means of combined immunocytochemistry and autoradiography, it was demonstrated that the majority (70-80%) of the tyrosine hydroxylase (TH)-immunoreactive cells that developed in the cultures possessed a transport system for NA. Catecholamine (CA) uptake also occurred in a small, but relatively constant, number of TH-negative cells, but was absent from substance P-containing neurons. In contrast to TH, which appears only after 3-4 d in vitro, cells capable of taking up NA with high affinity were found in DRG cultures after only a few hours, and a small number (less than 0.5% of the total cell population) was detected in freshly removed, uncultured ganglia. Such cells did not react with antibodies directed against substance P or neurofilament proteins. We conclude that autonomic precursors are identifiable in a subset of non-neuronal DRG cells, prior to full expression of a noradrenergic phenotype, by their possession of a high-affinity uptake system for CA.

Developmental capacities of avian embryonic dorsal root ganglion cells: neuropeptides and tyrosine hydroxylase in dissociated cell cultures.

Dorsal root ganglia (DRG) from quail embryos of 10-15 days of incubation (E10-15) contain a subpopulation of cells, distinct from postmitotic neurons, that can, under suitable conditions of culture in vitro, differentiate into neuron-like cells that display a variety of adrenergic properties, including tyrosine hydroxylase (TH) immunoreactivity (Xue et al., Proc. Natl. Acad. Sci. U.S.A., 82 (1985) 8800-8804). The present study was undertaken to determine whether other markers typical of autonomic sympathetic nerve cells are also expressed in the same system. Cells immunoreactive for vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) were found to differentiate continually from non-dividing precursors in all cultures of dissociated E10 quail DRG grown in the presence of chick embryo extract. Whereas VIP was already present (in a minute number of cells) in DRG in situ, NPY could not be detected before 3 days of culture, when it appeared simultaneously with TH. Double immunostaining experiments showed that most VIP-positive cells and about half the NPY-positive cells also displayed TH-immunoreactivity. On the other hand, there was no overlap between the substance P-containing neuronal population and any of the cells containing TH, NPY or VIP. These observations are pertinent to the problem of the segregation of autonomic and sensory cell lines during peripheral nervous system ontogeny.