Clinical Significance and Prognostic Value of the Expression of LAG-3 and FGL1 in Esophageal Squamous Cell Carcinoma.

In this retrospective study, we analyzed the expression of lymphocyte activating gene 3 (LAG-3) and fibrinogen-like protein 1 (FGP1) mRNA and the corresponding proteins in 78 patients with esophageal squamous cell carcinoma (ESCC) to evaluate the clinical significance and prognostic value. mRNA and protein expression were analyzed by reverse transcription PCR and Western blotting, respectively. The expression of LAG-3 and FGL1 mRNA and the corresponding proteins in tumor tissues were significantly increased in comparison with the normal esophageal mucosa. The overexpression of LAG-3 significantly correlated with the content of tumor-infiltrating lymphocytes (TILs), tumor differentiation, and TNM stage. The overexpression of FGL1 also significantly correlated with TILs, TNM stage, and lymph node metastasis. Kaplan-Meier survival analysis showed that tumor diameter, TNM stage, lymph node metastasis, LAG-3 and FGL1 protein expression were related to the progression-free survival (p<0.05). Multivariate Cox regression showed that the level of FGL1 and TNM stage were independent prognostic factors of progression-free survival. We speculated that the tumor microenvironment of ESCC induces immunosuppression due to up-regulated expression of LAG-3 and FGL1 in the tumor tissues, which promotes tumor growth.

[Clinical characteristics of 272 437 patients with different histopathological subtypes of primary esophageal malignant tumors].

Objective: To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT). Methods: A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results: A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment. Conclusion: ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Isolation of a Bacillus cereus strain HBL-AI and its application for production of Trans-4-hydroxy-l-proline.

A new trans-4-hydroxy-l-proline (trans-Hyp) producing Bacillus cereus HBL-AI, was isolated from the air, which was screened just using l-proline as carbon and energy sources. This strain exhibited 73·4% bioconversion rate from initial l-proline (3 g l-1 ) to trans-Hyp. By sequencing the genome of this bacterium, 6244 coding sequences were obtained. Genome annotation analysis and functional expression were used to identify the proline-4-hydroxylase (BP4H) in HBL-AI. This enzyme belonged to a family of 2-oxoglutarate-related dioxygenases, which required 2-oxoglutarate and O2 as co-substrates for the reaction. Homologous modelling indicated that the enzyme had two monomers and contained conserved motifs, which included a distorted 'jelly roll' ß strand core and the residues (HXDXnH and RXS). The engineering Escherichia coli 3 Δ W3110/pTrc99a-proba-bp4h was constructed using BP4H, which transformed glucose to trans-Hyp in one step with high concentration of 46·2 g l-1 . This strategy provides a green and efficient method for synthesis of trans-Hyp and thus has a great potential in industrial application.

Cordycepin inhibits cell growth and induces apoptosis in human cholangiocarcinoma.

The purpose of this study was to explore the role of cordycepin in human cholangiocarcinoma (CCA) cell growth and apoptosis. In the present study, colony formation assay, cell-counting kit-8 (CCK-8) assay and tumor xenograft experiment were performed to evaluate the effect of cordycepin on human CCA cell growth in vitro and in vivo; flow cytometric analysis was performed to evaluate the effect of cordycepin on cell apoptosis; quantitative real-time reverse transcription PCR (qRT-PCR) and western blot assays were performed to evaluate the expression levels of Caspase-3, Bcl-2 and Bax. The results showed that cordycepin inhibited cell growth in QBC939 and RBE cells in vitro and it could also inhibit QBC939 cells growth in vivo. Furthermore, the flow cytometric analysis, qRT-PCR and western blot assays showed that cordycepin could trigger QBC939 and RBE cells apoptosis by regulating the expression levels of Caspase-3, Bcl-2 and Bax. And we proposed that cordycepin could inhibit human CCA cell growth in vitro and in vivo, while, this function is related to the induction of cell apoptosis.

Long non-coding RNA MALAT1 promotes cholangiocarcinoma cell proliferation and invasion by activating PI3K/Akt pathway.

Increasing evidence indicated that metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acted as a key regulator in the proliferation and invasion of several cancers. However, the function of MALAT1 in the development of cholangiocarcinoma has not been experimentally established. In the present study, the expression levels of MALAT1 in cholangiocarcinoma cell lines were detected by quantitative real-time PCR. The effects of MALAT1 knockdown on the cell proliferation and invasion of cholangiocarcinoma cells were detected with Cell Counting Kit-8 (CCK-8), colony formation assay and Trans-well assay, respectively. The expressions of epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, Vimentin) were evaluated to discover whether the process of EMT was involved. We also evaluated the expression of phos-phatidylinositol-3-kinase/serine/threonine kinase (PI3K/Akt) signaling pathway proteins (PI3K, p-PI3K, Akt, p-Akt) to determine the associated molecular mechanism. And we discovered that MALAT1 was up-regulated in cholangiocarcinoma cancer cells. CCK-8, colony formation and trans-well assay showed that the proliferation and invasion of QBC-939 and RBE with MALAT1 knockdown were inhibited. Moreover, MALAT1 could promote EMT in cholangiocarcinoma cells. In addition, MALAT1 may activate PI3K/Akt pathway. These results indicated that MALAT1 promoted cholangiocarcinoma cell proliferation and invasion. The effects of MALAT1 on cholangiocarcinoma cells might be through activating the PI3K/Akt signaling pathway. These investigations may facilitate a better understanding of MALAT1 and it might be a potential therapeutic target for the treatment of cholangiocarcinoma.

Association of polymorphism in ICAM-1 (K469E) and cytology parameters in patients' initial blood test with acute ischemic stroke.

Acute ischemic stroke (AIS) has become a serious health problem in many countries because of its poor outcome and worsening epidemic trend. Early identification of genetic risk factors and physiological indicators for stroke occurrence may help to reduce the incidence of stroke. Therefore, we conducted a case-control study including 50 AIS patients and 50 healthy individuals from a Chinese population to explore the association between AIS and patient complete blood profiles and the association between AIS and the genetic polymorphism K469E in intercellular adhesion molecule-1 (ICAM-1). Compared to the control group, AIS patients showed a high percentage of mononuclear cells, low platelet count, low ratio of platelet to lymphocyte count, high frequency of the 469K allele, and low frequency of the 469E allele. White blood cell count, percentage of neutrophils, percentage of lymphatic cells, platelet distribution width, mean platelet volume, and platelet hematocrit levels showed no significant differences between the 2 groups and between different genotypes. Our results suggested an association of elevated levels of mononuclear cells and reduced platelet count with higher AIS risk. Our results also supported the hypothesis that the KK genotype at the K469E locus in ICAM-1 is a risk factor for AIS.

A Chinese patient with Madelung's disease and alcoholic cardiomyopathy: a case report and literature review.

BACKGROUND: Madelung's disease is a rare disorder characterized by massive deposits of excess subcutaneous fat around the neck, shoulders, arms, and other parts of the body. It has a high prevalence among middle-aged alcoholic men in Mediterranean countries and a low incidence in Asian populations. Although patients with Madelung's disease are often associated with a variety of alcohol-induced metabolic disorders, the comorbidity of alcoholic cardiomyopathy is rarely reported, probably because of its low incidence or neglect by clinicians. CASE REPORT: A 67-year-old man with a 10-year history of soft fat masses in the neck developed chest tightness and shortness of breath on exertion for the past 2 years. Laboratory tests revealed elevated γ-glutamyl transferase, glucose intolerance, hyperuricemia, hyperlipidemia, and anemia. Computed tomography of the neck showed symmetric nonencapsulated fat deposits, mainly in the anterior cervical regions. Echocardiography showed left heart enlargement and severe global left ventricular systolic dysfunction with an ejection fraction of 31%. Coronary angiography revealed 40-50% stenoses of the left anterior descending and right coronary arteries. After the exclusion of other causes of dilated cardiomyopathy, the patient was finally diagnosed with type I Madelung's disease and alcoholic cardiomyopathy. He underwent lifestyle changes, including reducing his alcohol intake, and received full pharmacological treatment for heart failure. One and a half years later, his cardiac function was partially restored, and all metabolic abnormalities improved except for elevated liver enzymes. CONCLUSIONS: Alcohol use disorder should be assessed in patients with newly diagnosed Madelung's disease. Screening for alcoholic cardiomyopathy in alcoholic patients with Madelung's disease is necessary for early detection of cardiac abnormalities and intervention to improve the prognosis of this group of patients.

An improved method assigning three-dimensional atomic potentials to multiple slices in exit-wave simulations of Transmission Electron Microscopy.

An atomic potential should be assigned to several slices in the multislice method owing to its three-dimensional (3D) distribution. Based on a formula including several Gaussian functions to fit electron atomic scattering factors, a simple analytical expression is proposed to calculate the potential of atoms projected onto multiple slices. The potential in 3D distribution is properly projected onto slices that do not contain the atomic centroid. Thus, the fluctuations in atomic altitude influence the assigning of atomic potentials is considered correctly. The projected potential is calculated in a reciprocal space and involves an accurate 3D atomic position. Tests conducted with a plausible Ag chain verify the good performance of this new approach. The simulated exit wave leaving a complex crystal of Ba6Nd2Ti4O17 demonstrates that the proposed simulation approach is better than the traditional multislice method.

RNA-sequencing reveals the metabolism regulation mechanism of sheep skeletal muscle under nutrition deprivation stress.

Although the skeletal muscle is one of the main sites of metabolism, little is known about the molecular mechanisms involving its response to nutrition stress. The aim of the study was to screen the transcriptome of sheep muscle to identify the metabolism-related genes under nutrition deprivation stress. Ten healthy adult female Small-tailed Han sheep with similar age and weight were randomly divided into a normal group and fasted group. After 3 days, three sheep were randomly selected from each group and the semitendinosus samples were subjected to RNA-sequencing (RNA-seq) and a series of analyses and function annotations. Compared with the normal group, 391 differentially expressed genes (DEGs) were identified in the fasted group that had obvious weight loss, including 278 down-regulated and 113 up-regulated genes. Gene Ontology enrichment annotation classified 228 DEGs in the metabolic process, 11 of which were new genes and only Sheep_newGene_4578 had been annotated by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The results of Clusters of Orthologous Groups annotation indicated that 11, 9, and 4 DEGs were respectively classified in lipid transport and metabolism, amino acid transport and metabolism, and carbohydrate transport and metabolism. In addition, KEGG enrichment analysis showed that there were not only pathways which were directly related to metabolisms such as protein digestion and absorption pathway, fatty acid metabolism pathway, and biosynthesis pathway of unsaturated fatty acids, but also PI3K-AKT pathway, AMPK pathway, MAPK pathway, and FoxO pathway which were important to metabolism among the top 20 pathways with the lowest significant Q value. The MCODE analysis of protein-protein interaction revealed that two identified subnetworks with top score were closely associated with metabolism. The correlation analysis showed that the mRNA levels of most of DEGs that might be related in the two subnetworks were significantly correlated respectively, and the mRNA levels of most of 10 metabolism-related DEGs including Sheep_newGene_4578 were significantly correlated. Finally, 16 random and 10 metabolism-related DEGs were chosen for confirmation by quantitative real-time PCR, demonstrating the same expression change as determined by RNA-seq. In conclusion, multiple interrelated metabolism-related DEGs in skeletal muscle contributed to the response of sheep to nutritional deprivation stress.

Pharmacokinetics, tissue distribution, and metabolism of novel DNA topoisomerase I inhibitor yuanhuacine in rabbit.

A liquid chromatography/tandem mass spectrometry method was developed and validated for the quantitative determination of yuanhuacine (YHC), a daphne diterpene ortho-ester anticancer agent, and identification of its metabolites. Pharmacokinetic behaviour, tissue distribution, and metabolism were investigated in rabbit. YHC plasma data best fitted to a two-compartment model and were characterized by an elimination half-life t(1/2)(beta) of 11.1 h following intravenous administration. Tissue distribution studies did not identify any tissues having a high affinity for YHC. The main metabolites are proposed to be M392I, M392II, and M390, resulting from the ortho-ester group and aromatic ester bond being cleaved off simultaneously during Phase I metabolism. This investigation contributes to an understanding of the metabolism of daphne diterpene ortho-esters.

Altered exosomal miR-181d and miR-30a related to the pathogenesis of CVB3 induced myocarditis by targeting SOCS3.

OBJECTIVE: MicroRNAs are a group of gene expression regulators and some of which have been confirmed to be associated with acute viral myocarditis (VM). This study aims to find new biomarkers for VM diagnosis and explore the roles of miRNAs during the pathogenesis of VM. PATIENTS AND METHODS: 23 patients with acute myocarditis and 12 controls were included in this research. The expression of 10 candidate miRNAs in the serum exosome was examined by qRT-PCR. The direct targets were predicted using bioinformatics tools and then confirmed by dual luciferase assay and immunoblotting. Levels IL-6 of cell culture supernatants were determined by enzyme-linked immunosorbent assay. Six weeks old male mice were injected intraperitoneally with Coxsackievirus B3 (CVB3) and then treated by miRNA inhibitors through tail vein injection. RESULTS: Five miRNAs were found to have disturbed expression in the exosome and may have the potential to be used as biomarker for VM diagnosis. Meanwhile, the expression of miR-30a and -181d was also altered in the cells after CVB3 infection. We identified SOCS3 as a direct target of miR-30a and -181d. Furthermore, during CVB3 infection, up-regulated miR-30a and -181d are related to enhanced IL-6 level via modulating SOCS3 expression. miRNA inhibitors injection increased mice survival rate after CVB3 infection. CONCLUSIONS: miR-30a and -181d contribute to the over-activated inflammatory response to viral infection of the heart during coxsackievirus infection.

Study on the relationship between miR-520g and the development of breast cancer.

OBJECTIVE: Breast cancer (BC) is one of the most common malignant tumors occurred in women. There is no sensitive and specific marker for early diagnosis, treatment and prognosis of breast cancer. It is suggested that miRNA may be a potential tumor marker for breast cancer. Mir-520g is considered to be associated with many tumors. This study aims to test the expression of mir-520g in peripheral blood of BC patients and healthy control. We also explored the relationship between mir-520g and several prognostic factors in breast cancer patients. PATIENTS AND METHODS: The peripheral blood of 86 cases with breast cancer (including 18 cases with stage 0, 24 cases of phase I, 20 cases of stage II, 24 cases of stage III) and 26 cases of healthy subjects were collected. The miR-520g level was measured by real-time quantitative PCR (RT qPCR) method. The correlation between plasma miR-520g level and the clinical stage, molecular subtype, receptors' expression and other factors related to the prognosis of the patients were examined. RESULTS: Plasma mir-520g expression levels were significantly higher in BC patients with lymph node metastatic and low differentiation degree grade (p = 0.033 and 0.016), and plasma miR-520g expression was significantly higher in breast cancer patients with mammary gland invasion (p < 0.01) and low expressed p53 (p = 0.0039). CONCLUSIONS: Highly expressed mir-520g is associated with lymph node metastasis and low differentiation of breast cancer, and also is associated with mammary gland invasion in breast cancer. This study suggests that mir-520g may be associated with some important prognostic factors in breast cancer patients, and may have a potential value for breast cancer marker.

Genomic organization of the human retinoblastoma gene.

Sequence analysis of the human retinoblastoma gene cDNA revealed the presence of repeated elements in the form of direct repeats, inverted repeats and dyad symmetries. The clustering of the dyad symmetrical elements in some exons, #16 and #17, coincides with the hot spots for structural aberrations of the RB-1 locus previously observed in tumors. The RB-1 gene is divided into at least 27 exons distributed over 200 kbp. Three potential Sp1 binding sites are presented within 600 bp upstream of the translation start site. A DNA fragment containing these Sp1 sites ligated to a promotorless CAT gene can promote its transcription in transfected cell culture.

The Rb gene suppresses the growth of normal cells.

The suppression of tumor formation, first demonstrated by somatic cell hybrid and microcell fusion experiments, suggests the existence of a class of genes that selectively suppress the growth of tumor cells but not normal cells. The reintroduction of these genes into tumor cells presumably renders the cells responsive to in vivo growth inhibitory environment. As the inheritance of a defective retinoblastoma gene (Rb-1) allele results in a predisposition to the development of various cancers, and since inactivation of both alleles are observed in tumor cells, the Rb gene has been suspected to have the ability to suppress tumor growth. Data presented here demonstrated that different types of normal cells, which have a limited life span, were also growth arrested by a transfected Rb gene. Cell lines which are resistant to the growth suppression effect of the Rb gene in vitro, retain the ability to form tumors in nude mice even in the presence of a stable and highly expressed wild type Rb protein. We conclude that while the Rb gene can suppress the growth of many tumor cell lines, its growth suppression effect is not tumor specific.

Hyperammonemia and anorexia in Morris hepatoma-bearing rats.

Inoculation of Buffalo rats with Morris hepatoma produced significant anorexia within four weeks and reduced body weight within two weeks. Blood ammonia concentration was increased by 113% when the rats were euthanized, five days after the development of anorexia. Infusing ammonium salts into normal Buffalo rats also induced anorexia at a blood ammonia concentration comparable to that observed in the tumor-bearing rats. Although ammonia-infused rats exhibited expected increases in brain tyrosine, tryptophan, and metabolites of dopamine and serotonin, these alterations were attenuated in the tumor-bearing rats. These results indicate that hyperammonemia may be a general consequence of experimental cancer and that the increase in ammonia concentration may be of primary importance in the development of experimental cancer-induced anorexia. The rather small alterations in neurotransmitter metabolism in anorectic tumor-bearing rats deemphasize the role aberrations in DA and 5-HT systems in the development of experimental cancer anorexia.

Free radicals-induced abnormal chondrocytes, matrix and mineralization.A new concept of Kaschin-Beck's disease.

Free radical generating substances, including fulvic acids, mycotoxins, Fe(II), etc attack chondrocytes, causing dedifferentiation. The dedifferentiated cells synthesize and secrete abnormal collagens rich in type I instead of the normal type II. These substances also directly attack collagens in the extracellular matrix. The abnormal collagens make the matrix change from hydroxyapatite crystallization inhibiting to promoting, and give crystals of low crystallinity, high aggregation and small size. This study leads to a new concept of the development of Kaschin-Beck's Disease (KBD).

[High myopia with esotropia fixus].

Most of the 19 cases of high myopia with esotropia fixus had the refractive error in childhood while esotropia fixus set in progressively during middle age. The eyeball was fixed in a position of marked adduction, and the forced duction test was positive. Electromyography showed normal innervation of the medial and lateral recti. Contracture of the medial rectus and thinning of the lateral rectus were observed during surgery. Histopathological examination revealed degeneration and fibrosis in the medial rectus. Etiology was not clear. Tenotomy of the medial rectus or that of the inferior rectus in addition, with fixation of the lateral rectus tendon to the lateral orbital rim produced satisfactory corrective results.

[Correction of post-operative strabismus after scleral encircling procedures].

The scleral encircling or explant procedures for treatment of retinal detachment usually lead to post-operative strabismus, due to traumatic muscular adhesion to the globe, explant interference with muscular function, nerve damage and muscular paralysis, excessive or prolonged stretching of the muscle and surgical muscular reattachment at an improper site. In addition, there may be muscular atrophy or cicatrizing transformation due to compression. In 3 patients, the authors made use of the cicatricial tissue in place of the original muscle with or without surgical adjustment of the other ocular muscles. Satisfactory cosmetic result and ocular motility were obtained in all 3 cases.

[Ultrastructural changes in the body wall and gut of Clonorchis sinensis in rats after albendazole treatment].

The effect of albendazole on the body wall and gut of Clornorchis sinensis was studied with transmission and scanning electron microscopes after albendazole administration to rats infected with Chonorchis sinensis at a single dose of 150 mg/kg. The results showed that swelling and adhesion of the projections of the tegument and gut microvilli occurred 1h after medication. Necrosis and disruption of the projections and the gut microvilli were seen at the 24th h. By the 72nd h, detachment of the partial projections were seen. The dynamic process of the damages observed on the tegument was identical with that of the gut microvilli (Figs. 1-10).