[Hsa_circ_0000670 promoted the progression of gastric cancer through the miR-515-5p/SIX1 molecular axis].

Objective: To explore whether hsa_circ_0000670 promotes the progression of gastric cancer by regulating the miR-515-5p/SIX1 molecular axis. Methods: The gastric cancer and adjacent normal tissues of 35 gastric cancer patients admitted to Rugao Hospital Affiliated to Nantong University from 2014 to 2015 were collected. The expression levels of circ_0000670, miR-515-5p and Sine oculis homeobox 1 (SIX1) in gastric cancer tissues and cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. The correlations between circ_0000670 and miR-515-5p, miR-515-5p and SIX1, circ_0000670 and SIX1 were analyzed by the Pearson method. Patients were divided into low circ_0000670 expression group (17 cases) and high circ_0000670 expression group (18 cases) based on the median of circ_0000670 expression level, and Kaplan-Meier was used to analyze the 5-year survival of patients. Cell proliferation was assessed via clone formation assay. Cell cycle and apoptosis were detected by flow cytometry. Wound healing and Transwell assays were used to detect cell migration and invasion ability. The targeting relationship between miR-515-5p and circ_0000670 or SIX1 was confirmed by the dual luciferase reporter assay. Nude mice were injected into HGC-27 cells transfected with sh-NC or sh-circ_0000670, and the volume and weight of the transplanted tumor were measured, also, the levels of circ_0000670, miR-515-5p and SIX1 in the transplanted tumor tissue were detected. Results: The expression levels of circ_0000670 and SIX1 in gastric cancer tissues and cell lines were significantly increased (P<0.05), while the expression levels of miR-515-5p were significantly decreased (P<0.05). The survival rate of patients in the low circ_0000670 expression group (82.4%) was significantly higher than that in the high circ_0000670 expression group (28.7%, P=0.034). Circ_0000670 was negatively correlated with miR-515-5p (r=-0.846, P<0.001), and miR-515-5p was negatively correlated with SIX1 (r=-0.615, P<0.001), but circ_0000670 was positively correlated with SIX1 (r=0.814, P<0.001). Transfection of si-circ_0000670 or miR-515-5p mimic could significantly reduce the number of clone-forming cells, migration distance, migration and invasion cells (P<0.05), and increase the ratio of G(0)/G(1) phase cells, apoptosis rate and the protein level of E-cadherin (P<0.05), decreased the proportion of S-phase cells and the protein level of Vimentin (P<0.05). The dual luciferase report assay confirmed that circ_0000670 could target miR-515-5p, and miR-515-5p could bind to SIX1. Co-transfection of si-circ_0000670 and miR-515-5p inhibitor could significantly attenuate the effects of si-circ_0000670 on cell proliferation, migration, invasion, cell cycle and apoptosis (P<0.05). Co-transfection of miR-515-5p mimic and pcDNA-SIX1 could significantly reduce the effects of miR-515-5p mimic on cell proliferation, migration, invasion, cell cycle and apoptosis (P<0.05). Compared with the sh-NC group [volume=(596.20±125.46) mm(3) and weight=(538.00±114.39) g], the volume and weight of transplanted tumors in the sh-circ_0000670 group [volume=(299.20±47.58) mm 3 and weight=(289.80±48.73 g)] were significantly reduced (P<0.05), the expression levels of circ_0000670 and SIX1 were significantly reduced (P<0.05), and the expression level of miR-515-5p was significantly increased (P<0.05). Conclusion: Knockdown of circ_0000670 could inhibit cell proliferation, migration, invasion of gastric cancer cells, induce cell cycle arrest in G(0)/G(1) phase and promote cell apoptosis by regulating the miR-515-5p/SIX1 axis.

Matrilin-3 alleviates extracellular matrix degradation of nucleus pulposus cells via induction of IL-1 receptor antagonist.

OBJECTIVE: The intervertebral disc contains abundant extracellular matrix (ECM) imbued with proteoglycans, collagens, and water. With the development of intervertebral disc degeneration (IVDD), the ECM undergoes changes characterized by loss of water content, proteoglycans, and collagen content. The purpose of this study was to explore the vital role of Matrilin-3, an ECM protein involved in the progress of IVDD. MATERIALS AND METHODS: NP cells were isolated from the patients' disc samples and exposed to recombinant human (rh)-Matrilin-3 protein (MATN3), and IL-1ß is used as a reducer of nucleus pulposus (NP) cells degeneration. Matrilin-3 and IL-1 receptor antagonist (IL-1Ra) were knocked down by siRNA transfection. Messenger RNA expressions of IL-1Ra, Collagen II, aggrecan, MMP-13, and ADAMTS-5 were determined using Real-Time quantitative Polymerase Chain Reaction (RT-qPCR). Later, the protein levels of IL-Ra, Collagen II, and aggrecan were also detected by Western blot. The IL-1Ra, MMP-13, and ADAMTS-5 dose of the supernatants in the culture medium was determined by enzyme linked immunosorbent assay (ELISA). Finally, immunofluorescence was used to expose the expression of Collagen II, aggrecan, and Collagen X. RESULTS: It was found that the expression of IL-1Ra was markedly increased in the present of MATN3 or IL-1ß, especially these two at once. Besides, MATN3 could upregulate Collagen II and aggrecan expressions, as well as inhibit the MMP-13 and Collagen X production of NP cells. However, the protective effects of Collagen II and aggrecan were abolished after Matrilin-3 silenced. Furthermore, IL-1ß downregulated the Collagen II and aggrecan but promoted the MMP-13 and Collagen X levels of NP cells, which were antagonized by the action of MATN3. Surprisingly, silencing of IL-1Ra significantly abolished the MATN3-induced the protective effects of ECM in NP cells. CONCLUSIONS: This study provides a novel viewpoint of Matrilin-3 in the ECM stability of NP due to its ability to activate IL-1Ra. It is considered that MATN3 efficiently protects ECM degeneration of human NP cells related to maintain the content of Collagen II and aggrecan, as well as inflammatory inhibition.

[Clinical analysis of gene mutation characteristics and mutation burden in myelodysplastic syndrome].

Objective: To investigate the relationship between gene mutation characteristics, mutation burden and general condition, disease subtype and karyotype of patients with myelodysplastic syndrome (MDS), and its clinical value. Methods: High-throughput sequencing was used to detect 65 blood tumor-related genes in 191 MDS patients and 9 secondary acute myelocytic leukemia patitents(SAML), and to analyze the characteristics of abnormal genes, mutation burden, as well as the relationship with disease subtypes, chromosome karyotypes and age. Results: Mutations were found in 148 patients (77.5%), including 47 abnormal genes and 186 mutation sites. And gene mutations were found in 9 SAML patients, the number of mutations was significantly higher than that in MDS patients (χ(2)=11.911, P=0.018). Among the abnormal genes, the mutation frequency of U2AF1 (37.3%) and ASXL1 (41.6%) were higher, and there were significant differences in mutation burden among different abnormal genes (F=91.946, P<0.001). There were differences in the number of gene mutations among different subtypes of MDS, and the number of EB-2 gene mutations was the highest (2.2±1.5). In SLD, MLD, EB-1 and EB-2, the proportion of carrying ≥ 3 mutations increased gradually (χ(2)=52.471, P=0.037). TP53 mutation was associated with abnormal karyotype (r(φ)=0.177, P=0.019), especially with complex karyotype (r(φ)=0.440, P<0.001), while NPM1 mutation is associated with normal karyotype (r(φ)=0.173, P=0.024). The number of mutations carried by patients under 30 years old was the least, and the number of mutations increased with the increase of age. The number of mutations was the most in patients aged 60 to 79 years old (P=0.017), and the mutation frequency of epigenetic related genes increased with the increase of age (P=0.041). Conclusions: The mutation characteristics and mutation load of MDS-related genes are closely related to clinical factors such as disease subtype, chromosome karyotype and patient age.

Peripheral T lymphocyte immunity and l-dopamine in patients with Parkinson's disease.

This study aimed to evaluate the efficacy of T-cell immune function and L-dopamine (L-DOPA) in patients with Parkinson’s disease (PD). Sixty subjects (included in the study group) with PD who were patients of the Neurology Department of The Affiliated Hospital of Hangzhou Normal University from July 2015 to March 2017 were selected. The study group was then categorized into groups according to the age of the patients, severity of disease, level of cognition, and treatment of L-DOPA. The control group (30 cases) was from the healthy population of the check-up center at The Affiliated Hospital of Hangzhou Normal University. The peripheral blood T-lymphocyte subsets of the study group were measured by direct immunofluorescence flow cytometry staining and compared with the control group. At the same time, correlation analysis was carried out on patients with different degrees of disease severity according to staging, different accompanying symptoms, and whether L-DOPA was administered. The results of the study show that the levels of CD4+, CD8+, CD3+, and CD4+/CD8+ peripheral blood in PD patients were significantly lower than those in the control group (P less than 0.05). It was found that the levels of CD4+, CD8+, CD3+, and CD4+/CD8+ decreased with age. The CD4+, CD8+, CD3+, and CD4+/CD8+ in patients with advanced stage PD were more significant than those with low PD stages (P less than 0.05). The levels of CD4+, CD8+, CD3+, and CD4+/CD8+ in the dementia group were significantly lower than those in the non-dementia group (P less than 0.05). The levels of CD4+, CD8+, CD3+, and CD4+/CD8+ in PD patients treated with L-DOPA were higher than those of PD patients without L-DOPA treatment (P less than 0.05). In conclusion, the immune function of T cells in patients suffering from PD is low, and the immune function of T cells in patients with severe disease is lower. Therefore, it is of certain significance to further study the pathophysiological mechanism of PD.

[Comparison of calcar replacement arthroplasty and Intertan nail in treatment of intertrochanteric fracture in the aged].

OBJECTIVE: To compare the clinical efficacy between calcar replacement arthroplasty and InterTan nail in treatment of intertrochanteric fractures in the aged. METHODS: From January 2008 to July 2013, a total of 58 elderly patients with intertrochanterie fracture , had been treated in Department of Orthopaedics, Tianjin Occupational Diseases Control and Treatment Hospital, were retrospectively compared.Half of them were treated with calcar replacement arthroplasty and half with InterTan nails.In the calcar replacement arthroplasty group, there were 8 men and 21 women, with an average age of (85.3±4.9) years, there were 8 A1, 16 A2 and 5 A3 fractures according to the AO classification.In the InterTan nail group, there were 10 men and 19 women, with an average age of (86.1±5.6) years, there were 11 A1, 15 A2 and 3 A3 fractures according to the AO classification.The 2 groups were compared in terms of the duration of the operation, intraoperative blood loss, hospitalization time, time of off-bed activity, complications, mortality rate, Harris hip score, short form 36 questionnaire score and ADL score. RESULTS: Twenty-seven patients in the calcar replacement arthroplasty group and twenty-six patients in the patients in each group were followed up InterTan nail group were follow-up for a period of 24 months.There were no significant differences between the 2 groups regarding hospitalization time, complications, mortality rate, Harris hip score, short form 36 questionnaire score and ADL score(P>0.05). However, the operative time, intraoperative blood loss, and the time of off-bed activity for the calcar replacement arthroplasty and InterTan nail groups was respectively (78±29) minutes vs (59±32) minutes, (316±185) ml vs (108±97) ml, and (15±5.8) days vs (32±12.7) days, all of them with a significant difierence(P<0.05). CONCLUSION: The patients in the calcar replacement arthroplasty group suffered longer operative time and more blood loss, but they can obtain early off-bed activity.The hospitalization time, complications, mortality rate, hip function and quality of Life between the two were the same.

The classification of posterior petrous meningiomas and its clinical significance.

This retrospective analysis of the clinical records of 42 patients was used to study the clinical significance of a classification system for posterior petrous meningiomas. According to clinical manifestations and radiological images, posterior petrous meningiomas were classified into three types: type I (cerebella type; 12 patients), tumours involved and compressed the cerebellum; type II (cranial nerve type; 16 patients), tumours involved the cranial nerves; and type III (combined type; 14 patients), tumours involved more than one structure such as the cerebellum, cranial nerves and the brain stem. All patients underwent microneurosurgery and the total resection rate was 90%. It was more difficult totally to resect type II and III tumours than type I tumours and the post-operative functional outcomes were worse. Microneurosurgical techniques and skills are critical to increase the total resection rate of posterior petrous meningiomas in order to decrease the mortality and disability rates.

Detection of cancer stem cells from the C6 glioma cell line.

Various malignant cancers have been found to contain a sub-population of stem cell-like tumour cells, or cancer stem cells (CSCs), however, culture methods for CSCs and the size of the fraction of CSCs in C6, which is a commonly used glioma cell line, remain controversial. In this study, we demonstrated that the C6 cell line contains a fraction of tumour cells that can form tumour spheres in a simplified serum-free neural stem cell medium and express CD133 and nestin, which are widely-used markers for brain CSCs. Immunohistochemistry and immunofluorescence confirmed the existence of CSCs both in the C6 cell line and C6 xenografts. Flow cytometry demonstrated that 4.02% of cells in the C6 cell line and 4.21% in the C6 xenografts presented as CSCs. These results confirm the fraction of CSCs in the C6 cell line and provide a simple and effective method for isolation of CSCs to study the initiation and progression of human glioma and, possibly, other malignant tumours.

Genetic polymorphisms in glutathione S-transferases T1, M1 and P1 and susceptibility to reflux esophagitis.

Recent studies indicate that the prevalence of reflux esophagitis (RE) in China is increasing. RE is one of the most common esophageal complications associated with gastroesophageal reflux disease (GERD) and RE-Barrett's esophagus-esophageal adenocarcinoma (EAC) sequence has been considered as an histogenesis model for EAC in Western countries. RE is only present in a subset of patients with GERD, suggesting an altered susceptibility to RE may exist in these GERD individuals. However, the genetic changes related with high susceptibility to RE is largely unknown. The polymorphisms in glutathione S-transferases (GSTs) T1, M1 and P1 have been reported with high susceptibity to esophageal cancer in Chinese people. The present case-control study was thus undertaken to characterize the genetic polymorphisms of GSTs and their correlation with susceptibility to RE. One hundred and nine patients with RE, 97 patients with nonerosive reflux disease (NERD) and 97 normal controls were recruited in this study. All the subjects were from Beijing, China, and received endoscopic examination and questionnaires for RE. Genomic DNA was extracted from the lymphocytes of peripheral blood for each subject. Genotypes of the GSTM1 and GSTT1 genes were analyzed by a multiplex PCR method. A-->G polymorphism of codon 104 of the GSTP1 gene was detected using PCR-based restriction fragment length polymorphisms (RFLP). The variant GSTP1 genotypes (*A/*Bomicron*B/*B) was found with a high frequency in the case with RE (40%), and followed by NERD (25%) and normal control (22%). The differences were statistically significant (P < 0.05). The risk for RE increased 2.42-fold [odds ratio (OR); 95% confidence interval (95% CI), 2.42 (1.22-4.80)] in the subjects with variant GSTP1 genotype. The subjects with positive variant GSTP1 genotypes and negative H. pylori infection showed increasing tendency for risk of RE [OR (95% CI), 2.67 (1.06-6.70)]. However, the subjects with GSTT1 and GSTM1 polymorphisms did not show any correlation with high risk for RE or NERD. No significant interactions were identified between the variant GSTs and cigarette smoking, or alcohol drinking and subtype of RE. The present result suggests that GSTP1 genetic polymorphism may be one of the high susceptibility factors involved in the mechanisms of RE. H. pylori infection may play a protective role against RE.

[Study on immunomodulating mechanism of the active fraction of liuwei dihuang decoction].

OBJECTIVE: To investigate the immunomodulating mechanism of 3A, an active fraction extracted from the ethanol-soluble portion of Liuwei Dihuang Decoction (LWDHD). METHODS: Influence of 3A on T-lymphocyte subsets, IgG production of B cells, interferon-gamma (IFN-gamma) mRNA expression in cyclophosphamide (Cy)-treated mice, tumor bearing mice and senescence accelerated mouse-prone/8 (SAMP8) were assessed in vivo and in vitro. RESULTS: Oral administration of 3A could elevate the decreased T-lymphocyte subsets in all model mice, it also could promote the IgG secretion of splenic cells in SAMP8 and increase IFN-gamma mRNA expression in Cy-treated mice. CONCLUSION: The immunomodulating effect of 3A is related with its effects in regulating ratio of T and B cells or in improving their functions.

[High-performance dynamic ECG review & analysis system].

This paper describes the design of a high-performance dynamic ECG Review & Analysis system used for not only clinical diagnosis but also medical research. A new ECG pattern classifier with Variable Area Template is applied to improve the quantity of ECG analysis results, while the optimized software design ensures the other performances. The results show that this system has higher performance than the old ones and is valuable for medical research.

[Effect of membrane modification by PEG on prolongation of circulation time of liposomes in blood in vivo].

PEG-PE (polyethylene glycol-phosphatidyl ethanolamine) of different molecular weight (2000 and 5000) were used to modify the membrane of liposomes. Large unilamellar liposomes containing PEG-PE were prepared by reversed phase evaporation. Fluorescent label-calcein was encapsulated at the internal water phase. To compare the differance between the modified and unmodified membrane, the stability in vitro and distribution in vivo were investigated. The results indicated that the circulation half-life for liposomes unmodified, modified by PEG (2000)-PE and modified by PEG (5000)-PE were 13, 21 and 75 (min) respectively. At 6 h after injection, the ratio b/R (b: distribution in blood, R: distribution in liver and spleen) were 0, 0.8 and 1.4, respectively. The results mean that the stability increased and circulation time was prolonged by the PEG-PE modified membrane. The effect of PEG-PE on membrane was found to be directly proportional to the chain length of the polymer.

[Application of western blotting technique in the serodiagnosis of Mycobacterium tuberculosis infection].

We have used western blotting technique for examining antibodies in patients with tuberculosis and in healthy individuals. In patients with tuberculosis, the number and intensity of the developed antigen bands were much higher than in normal individuals. Furthermore, disease-associated protein antigens of 30,000-43,000, 28,000 and 25,000 were found to react with patients' sera. A specificity of 0.9639 and sensitivity of 0.9012 were obtained about 30,000-43,000 antigens. 25,000 antigen reacted with 41.74% of patients' sera but with none of the controls.

Single-sweep polarographic investigation of concanavalin A and its interaction with polysaccharide.

A simple, rapid, and sensitive single-sweep polarographic method has been developed for investigation of concanavalin A (con A) and its interaction with selected polysaccharides. In a solution containing 0.001 M 2,2'-bipyridine, 0.015 M hexamethylenetetramine, and 0.1 M sodium chloride, con A exhibits a single-sweep polarographic wave, and the cathodic peak potential is -1.50 V (vs SCE). The peak current varies linearly with con A concentration over a range of 1.0 x 10(-8) to 1.2 x 10(-7) M by derivative single sweep polarography. A preliminary discussion on properties of the con A polarographic wave has been made. In addition, it has been demonstrated that single sweep polarography can be a useful method for studies on interactions of con A with its complementary polysaccharides in solution.

[Effects of metoclopramide on slow response action potentials of myocardium].

The effects of metoclopramide (Met) on the action potential of rabbit sinus node cells and slow response action potential of guinea pig papillary muscles were studied with the intracellular microelectrodes. Met 10 mumol.L-1 prolonged the action potential duration at 90% repolarization (APD90) of SA node cell and sinus cycle length (SCL). Met 100 mumol.L-1 caused a decrease in action potential amplitude (APA), Vmax and the slope of phase 4 of action potential of SA node cell. APD90 and SCL were prolonged further. For slow response action potential induced by KCl (25 mmol.L-1), Met 100 mumol.L-1 produced a decrease in APA and depolarization rate. Met 10 mumol.L-1 began to suppress the spontaneous electrical activities induced by barium ion. These findings suggest that Met probably has the effect of blocking calcium inward current in myocardium.

[Effects of metoclopramide on Vmax of action potentials in guinea pig papillary muscles].

The effects of metoclopramide (MCP) on Vmax of action potentials of guinea pig papillary muscles were studied with intracellular microelectrodes. MCP 10-550 mumol/L abbreviated the action potential duration at 90% repolarization (APD90) and decreased the maximal rate of rise of action potential (Vmax) dose-dependently. MCP caused rate-dependent and voltage-dependent reductions of Vmax and retardation of the recovery of Vmax. The results present the possible causes of depressant effect of MCP on Na channel.

Systemic mycoses in the United States, 1980-1982.

To evaluate temporal trends in the incidence of systemic mycoses resulting in hospitalization, we reviewed hospital discharge data from the Professional Activity Study of the Commission on Professional and Hospital Activities for 1980-1982. In comparison with results from similar studies in 1970 and 1976, the incidences of the mycoses that are more likely to be opportunistic and/or hospital-acquired infections (candidiasis, aspergillosis, cryptococcosis) increased in 1980-1982 as did the incidences of sporotrichosis and actinomycosis. At the same time the incidences of most community-acquired mycoses (histoplasmosis, coccidioidomycosis, and blastomycosis) remained stable or declined. These results are consistent with the increasing number of immunocompromised patients with severe underlying illnesses, who are at risk of developing opportunistic mycoses.