Abstract Polysaccharides are a class of biological macromolecules made up of sugar chains with multiple glycosidic bonds. To explore the extracted, purified, carboxymethylated and phosphorylated modified to obtained carboxymethylated polysaccharides (HSPS) and phosphorylated polysaccharides (HSPP), and the HSPS and HSPP were characterized and analyzed, and their antioxidant activity measured. Response surface design was used for the synthesis process of HSPS and HSPP, the structure characterization and antioxidant activities of HSPS and HSPP were evaluated. The results showed that the Mw and Mn of HSP were 14.66 kDa and 8.39 kDa, respectively. The monosaccharides were mainly galactose, glucose, xylose, and arabinose in a molar ratio of 37.68%: 24.43%: 13.63%: 9.18%. The optimal synthesis conditions for HSPS were as follows: reaction time of 2.01 h, reaction temperature of 44.61°C and the amount of monochloroacetic acid necessary was 1.18 g. The optimal synthesis conditions for HSPP were: reaction time of 4.28 h, reaction temperature of 80.78°C, and reaction pH of 8.98. HSPS and HSPP have scavenging effects on ABTS, super oxideanion and hydroxyl radicals. The HSPS and HSPP were characterized and analyzed, and their antioxidant activity measured.

Abstract Background: UVB irradiation can cause acute damage such as sunburn, or photoaging and melanoma, all of which are major health threats. Objective: This study was designed to investigate the mechanism of skin photoaging induced by UVB radiation in mice through the analysis of the differential expression of miRNAs. Methods: A UVB irradiation photoaging model was constructed. HE and Masson special stains were used to examine the modifications in the epidermis and dermis of mice. The miRNA expression profiles of the mouse skin model exposed to UVB radiation and the normal skin of mice were analyzed using miRNA-sequence analysis. GO and Pathway analysis were employed for the prediction of miRNA targets. Results: A total of 23 miRNAs were evaluated for significantly different expressions in comparison to normal skin. Among them, 7 miRNAs were up-regulated and 16 were down-regulated in the skin with photoaging of mice exposed to UVB irradiation. The differential expression of miRNA is related to a variety of signal transduction pathways, among which mmu-miR-195a-5p and mitogen-activated protein kinase (MAPK) signal pathways are crucial. There was a significant differential expression of miRNA in the skin of normal mice in comparison with the skin with photoaging induced by UVB irradiation. Study limitations: Due to time and energy constraints, the specific protein level verification, MAPK pathway exploration, and miR-195a-5p downstream molecular mechanism need to be further studied in the future. Conclusions: UVB-induced skin photoaging can be diagnosed and treated using miRNA.

Abstract A response surface method (RSM) was employed to optimize polysaccharide extraction from Panax japonicus (PJPS). Also, carboxymethyl substitution of PJPS (CM-PJPS) was studied using a response surface methodology. A three-variable Box–Behnken design (BBD) methodology was applied. RSM analyses revealed that conditions that maximized polysaccharide yield were as follows: a liquid-to-material ratio 22:1, an extraction temperature of 83.99 °C and an extraction duration of 2.32 h. Under optimum extraction conditions, the average absorbance value of PJPS was 0.6001, and the purity of PJPS was 98.2%. A quadratic regression model for CM-PJPS was obtained via BBD. The optimum extraction conditions for the process were determined to include a 1.38 h reaction time, 1.24 g monochloroacetic acid (MCA) and a reaction temperature of 52.85 °C. Under optimum extraction conditions, the average degree of CM-PJPS substitution was 0.9733. The structure of the PJPS was examined by means of Fourier transform infrared spectroscopy (FTIR). CM-PJPS exhibited the strongest anti-oxidant activity in vitro. In addition, CM-PJPS inhibited Skov3 and A2780 cancer cell proliferation. Our data revealed that CM-PJPS is a promising natural antioxidant with potential value as a food supplement and for treatment of cancer.

Abstract In this study, we extracted lotus root polysaccharide (LRP) and synthesised phosphorylated lotus root polysaccharide (PLP) using response surface methodology (RSM). RSM analyses revealed that the optimal conditions for PLP synthesis were a reaction duration of 7 h, temperature of 70 °C and pH of 11.38. Under these conditions, the predicted degree of substitution (DS%) was determined to be 9.96%. The structure of the LRP1 was examined by ultraviolet (UV) spectroscopy scan, Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) (1H and 13C). The monosaccharide composition of LRP1 was determined to be mannose (0.12%), ribose(0.18%), glucuronic acid(0.60%), galacturonic acid(0.09%), glucose(98.79%) and galactose(0.21%). The number average molecular weight (Mn) and the weight average molecular weight (Mw) of LRP1 were 10236 and 251783 g/mol. LRP1 exhibited high antioxidant activities in scavenging ABTS radicals, Superoxide anion radicals and Metal ion scavenging activity. PLP exhibited strong antioxidant activity in vitro. In addition, PLP inhibited Skov3 cancer cell proliferation and induced reactive oxygen species (ROS) production. Our data revealed that PLP is a promising natural antioxidant with potential value as a food supplement and for the treatment of cancer.

Abstract In this study, lotus root polysaccharide was taken as raw material. The preparation technology of its polysaccharide selenium was established by response surface design, and its antioxidant activity in vitro was studied. Selenium content was taken as the detection index, and the effects of reaction time, the ratio of sodium selenite to the polysaccharide, and the volume fraction of nitric acid on selenium content were investigated. Through the response surface method, the optimal synthesis conditions of lotus root polysaccharide selenium were obtained as follows: reaction time of 5.36h, the ratio of sodium selenite and lotus root polysaccharide is 0.78, the volume fraction of nitric acid 0.47%, the selenium content of lotus root polysaccharide selenium 0.1106%. Selenium, a polysaccharide from lotus root, had a scavenging effect on ABTS radical and superoxide ion, and the maximum scavenging rate was 52.17% and 85.23%, respectively. Aa shows that the synthesized lotus root polysaccharide selenium can be used as an antioxidant or functional food.

Abstract In this study, we extracted lotus root polysaccharide (LRP) and synthesized carboxymethylated lotus root polysaccharide (CM-LRP) using response surface methodology (RSM). The monosaccharide component of LRP was analyzed by pre-column derivatization high performance liquid chromatography (PCD-HPLC). The polysaccharide was characterized by ultraviolet (UV) spectroscopy scan, Fourier transform infrared spectroscopy (FTIR), circular dichroism spectrometer (CD), and scanning electron microscopy (SEM). It was found that the main monosaccharide component of LRP was glucose, accounting for more than 98% of the total polysaccharide component. RSM analyses revealed that the optimal conditions for CM-LRP synthesis were a reaction duration of 2.22 h, the chloroacetic acid dosage is 2.02 g, and a temperature of 46.87 °C. Under these conditions, the predicted degree of substitution (DS) was determined to be 0.5101. The superoxide anion, ferrous ion and hydroxyl radical antioxidant system were constructed to study the antioxidant activity of LRP and CMLRP. The activity of CMLRP to remove ferrous ions and hydroxyl radicals were significantly stronger than that of LRP, and it also showed a certain concentration-dependent. LRP has a better scavenging ability than CMLRP in scavenging superoxide anion free radicals. Our data revealed that CM-LRP is a promising natural antioxidant with potential value as a food supplement.

Abstract Objective: To evaluate the mid-term survival rate after tricuspid valve replacement (TVR). Methods: We retrospectively studied 110 consecutive patients who underwent TVR from January 2007 to November 2017. A survival analysis was performed with the Kaplan-Meier method and the log-rank test. Results: The median survival was 65.81 months. Mean age was 50 (range 39 to 59) years. Forty-eight patients (43.6%) were male, and 62 patients (56.4%) were female. Most of the patients (78.5%) were categorized into the New York Heart Association (NYHA) functional classes III/IV. Seventy-two patients (65.5%) had isolated TVR. Six-three patients (57.3%) had previously undergone heart surgery. The Kaplan-Meier survival rates at one year, three years, and five years were 59.0%±5%, 52.0%±6%, and 48.0%±6%, respectively. A Cox regression analysis demonstrated that the risk factors for mid-term mortality were advanced NYHA class (hazard ratio [HR] 2.430, 95% confidence interval [CI] 1.099-5.375, P=0.028), need for continuous renal replacement therapy (CRRT) treatment (HR 3.121, 95% CI 1.610-6.050, P=0.001), and need for intra-aortic balloon pump (IABP) treatment (HR 3.356, 95% CI 1.072-10.504, P=0.038). Conclusion: In TVR, impaired cardiac function before the operation and a need for CRRT or IABP treatment after the operation is independently associated with increased mid-term mortality.

The aim of this study was to perform an updated meta-analysis to quantitatively investigate the association between G20210A polymorphism of Prothrombin gene and the risk of retinal vein occlusion (RVO), based on the available publications with inconsistent results. We utilized the Stata software to perform the heterogeneity test, association test, Begg's and Egger's tests, and sensitivity analysis. We searched three on-line databases (PubMed, Embase, and WOS) and obtained a total of 422 articles. Based on our selection criteria, 24 case-control studies were finally enrolled in this overall meta-analysis; a subgroup analysis by the factors ethnicity, control source, and RVO type was done. Through the association test of overall meta-analysis, we did not observe a significant difference between RVO cases and controls under the A vs G (allele) (z=1.49, P=0.137), A vs G (carrier) (z=1.42, P =0.155), GA vs GG (z=1.50, P=0.135), and GA+AA vs GG (z=1.50, P=0.135). Furthermore, we observed similar negative results in the association test of subgroup analysis (all P>0.05). Heterogeneity, Begg's, and Egger's tests excluded the presence of high heterogeneity and publication bias. Statistically stable results were observed in the sensitivity analyses. Based on integrated analysis of the current evidence, Prothrombin gene G20210A polymorphism is likely unrelated to the risk of RVO.

Abstract Purpose: To establish a method for the preparation of zoledronate liposome and to observe its effect on inducing the apoptosis of rat liver Kupffer cells. Methods: Zoledronate was encapsulated in liposomes, and then the entrapment rate was detected on a spectrophotometer. The prepared Zoledronate liposome (0.01 mg/mL) was injected into the tail vein of SD rats. Three days later, the number of Kupffer cells (CD68 positive) in rat liver tissue was detected by immunohistochemistry. Flow cytometry was used to detect the apoptosis rate of the isolated liver Kupffer cell cultured in vitro. Results: The entrapment rate of Zoledronate was 43.4±7.8%. Immunohistochemistry revealed that the number of Kupffer cells was 19.3±2.1 in PBS group and 5.5±1.7 in Zoledronate liposome group, with a significant difference (P<0.05). The apoptosis rate of Kupffer cells was 4.1±0.8% in PBS group, while it was 9±2.2% and 23.3±5.9% in Zoledronate liposomes groups with different concentrations of Zoledronate liposome (P<0.05). Conclusions: Zoledronate liposomes can effectively induce the apoptosis of Kupffer cells in vivo and in vitro, and the apoptosis rate is related to the concentration of Zoledronate liposome. To establish a rat liver Kupffer cell apoptosis model can provide a new means for further study on Kupffer cell function.

Abstract Background: The prevalence of obesity is increasing worldwide. Oxidative stress plays an etiological role in a variety of obesity-related metabolic disorders. 4-hydroxynonenal (4-HNE) is the most abundant and reactive aldehydic product derived from the peroxidation of n-6 polyunsaturated fatty acids with diverse biological effects that are not well detailed. Obesity is associated with decreased plasma adiponectin concentrations and increased production of lipid peroxidation products, including 4-HNE, in adipose tissue. There may be some association between the level of adipokines and 4-HNE. Material and methods: To analyze the associations between 4-HNE and classical adipokines, namely, adiponectin and leptin in a Chinese population, the plasma 4-HNE, adiponectin and leptin levels of 160 non-diabetic obese (NDO) patients and 160 healthy subjects were determined by ELISA, and their associations with adiposity, glucose, lipid profiles, insulin secretion and insulin sensitivity were studied. Results: Plasma 4-HNE levels were significantly increased in patients with NDO compared with healthy controls (p < 0.01). 4-HNE was negatively correlated with adiponectin and positively correlated with leptin. The plasma levels of 4-HNE were significantly correlated to several parameters involved in body mass index (BMI) and insulin resistance (IR). The 4-HNE levels were positively correlated with BMI and negatively correlated with insulin sensitivity. Conclusion: We conclude that 4-HNE is associated with the secretion of adiponectin and leptin and is correlated with IR in NDO humans. These findings indicate a pro-inflammatory role of 4-HNE in NDO patients, which supports the potential role of 4-HNE in the development of obesity-related disorders.

Resumen Introducción: la prevalencia de la obesidad está aumentando en todo el mundo. El estrés oxidativo desempeña un papel etiológico en una variedad de desórdenes metabólicos relacionados con la obesidad. El hydroxynonenal 4 (4-HNE) es el aldehido más abundante y más reactivo derivado de la peroxidación de los ácidos grasos poliinsaturados n-6, con efectos biológicos diversos que no son bien conocidos. La obesidad se asocia a concentraciones disminuidas de adiponectinas en el plasma y a un aumento en los productos de la peroxidación lipídica, incluyendo el 4-HNE, en tejido adiposo. Puede haber una cierta asociación entre el nivel de adipoquinas y el 4-HNE. Material y métodos: para analizar las asociaciones entre 4-HNE y las adipoquinas clásicas, adiponectina y leptina se determinaron por ELISA los niveles de adiponectina y de leptina, así como de 4-HNE, en una población de 160 pacientes chinos obesos no diabéticos (NDO) y de 160 controles sanos, y se estudió su asociación con adiposidad, perfil glucémico y lipídico, secreción de la insulina y sensibilidad de la insulina. Resultados: los niveles de 4-HNE aumentaron significativamente en los pacientes con NDO comparado con los controles sanos (p < 0,01). El nivel de 4-HNE se correlacionó negativamente con la adiponectina y positivamente con la leptina. Los niveles de 4-HNE se correlacionan positivamente con el IMC y negativamente con la sensibilidad a la insulina. Conclusión: concluimos que el 4-HNE está asociado a la secreción de adiponectina y de leptina y correlacionado con la resistencia a la insulina en sujetos obesos no diabéticos. Estos resultados indican un papel proinflamatorio del 4-HNE en pacientes NDO, que apoya el papel potencial del 4-HNE en el desarrollo de alteraciones relacionadas con la obesidad.

RESUMO CONTEXTO: Leucemia promielocítica aguda (LPA) compreende 8% a 10% dos casos de leucemia mieloide aguda (LMA). A remissão em casos de LPA de alto risco ainda é dificilmente conseguida, e recorrência é comum. RELATO DE CASO: Descrevemos aqui um caso de LPA com glóbulos brancos elevados no exame de sangue e a morfologia variante hipogranular na medula óssea, juntamente com fms-like tirosina-quinase-3 com mutações de duplicação em tandem interna (FLT3-ITD) e a isoforma bcr-3 de PML- RARα. Mais importante, detectamos alto nível de gene do tumor de Wilms (WT1) em blastos medulares por RT-PCR (reverse transcription polimerase chain reaction). Até agora, não há conclusões claras sobre a associação entre os níveis de expressão WT1 e APL. Este paciente recebeu sucessivamente regime de tratamento combinado, de hidroxicarbamida, trióxido de arsênico e idarrubicina e citarabina, alcançando finalmente a remissão completa. Infelizmente, em seguida, ele morreu de repente de hemoptise maciça devido a uma infecção pulmonar. CONCLUSÃO: Com base em nossos resultados e numa revisão da literatura, a função anormal de WT1 pode ser um fator de alto risco em casos de APL. Novos estudos, com o objetivo de avaliar o impacto da expressão de WT1 no prognóstico dos doentes com APL, são de interesse.

ABSTRACT CONTEXT: Acute promyelocytic leukemia (APL) accounts for 8% to 10% of cases of acute myeloid leukemia (AML). Remission in cases of high-risk APL is still difficult to achieve, and relapses occur readily. CASE REPORT: Here, we describe a case of APL with high white blood cell counts in blood tests and hypogranular variant morphology in bone marrow, together with fms-like tyrosine kinase-3 with internal tandem duplication mutations (FLT3-ITD), and bcr-3 isoform of PML-RARα. Most importantly, we detected high level of Wilms’ tumor gene (WT1) in marrow blasts, through the reverse transcription polymerase chain reaction (RT-PCR). To date, no clear conclusions about an association between WT1 expression levels and APL have been reached. This patient successively received a combined treatment regimen consisting of hydroxycarbamide, arsenic trioxide and idarubicin plus cytarabine, which ultimately enabled complete remission. Unfortunately, he subsequently died of sudden massive hemoptysis because of pulmonary infection. CONCLUSION: Based on our findings and a review of the literature, abnormal functioning of WT1 may be a high-risk factor in cases of APL. Further studies aimed towards evaluating the impact of WT1 expression on the prognosis for APL patients are of interest.

PURPOSE: To observe the effect of cholecystectomy on the changes of motion pattern of Beagle dogs' sphincter of Oddi (SO), and investigate the modulatory role of nitric oxide (NO) and cholecystokinin (CCK) in the regulation of SO.METHODS: Pressure of common bile duct, SO motility, response to bolus injections of cholecystokinin (CCK, 20 ng/kg and 100 ng/kg), basal pressure (BP) and phasic contraction amplitude (PCA) were measured respectively by manometry in six Beagle dogs before and after cholecystectomy.RESULTS: After cholecystectomy, the pressure and diameter of common bile ducts (CBD) was significantly increased (p<0.01); BP and phasic contraction frequency (PCF) were also increased, however, no significant differences were found between the two groups; the SO motilities was not significantly changed. The relaxation responded to physiological dose of CCK (20ng/kg) was decreased, while bolus-dose of CCK (100ng/kg) induced rapid contractions and decreased PCA after cholecystectomy. The regulation pattern of SO pressure modulated by NO and its inhibitor had changed after cholecystectomy.CONCLUSION: After cholecystectomy in Beagle dogs, no obviously change of motion pattern of SO was observed through self-compensation, but these compensations may lead to some changes of regulation pattern of CCK and NO on SO.

PURPOSE: Natural orifice transluminal endoscopic surgery (NOTES) is a new technique. This study describes our initial experience of NOTES and investigates the feasibility of transumbilical endoscopic cholecystectomy (TUEC). METHODS: Eight domestic pigs were submitted to TUEC. After establishment of pneumoperitoneum, a bi-channel endoscope was placed through an infra-umbilical trocar. The gallbladder fundus was lifted by a grasper. The cystic duct and artery was dissected with a flexible hook and clipped by a clip fixing device. The specimen was extracted through the infra-umbilical trocar. RESULTS: The mean operation time was 114 minutes, ranging from 75 to 155 minutes. All the gallbladders were removed successfully. There was one case of subtotal resection, two cases of bleeding and three cases of bile leakage. CONCLUSION: Transumbilical endoscopic cholecystectomy is feasible although it needs more support of experiments and techniques before being applied on human subjects.

The aim was to understand the anatomical features of the venous valve in Macaca fascicularis and to compare it with that of humans. The bilateral lower limbs (24 limbs from 12 animals) of Macaca fascicularis cadavers were dissected, and the femoral veins (FVs) were equally divided into distal, intermediate, and proximal sections. The external diameter of the FV in each section was measured. The venous valves were observed microscopically and stained with hematoxylin and eosin as well as trichrome. Data describing the human venous valve were collected from the current literature. No great saphenous veins were found among the 24 lower limbs from the Macaca fascicularis cadavers. The external diameters of the FVs in the distal, intermediate, and proximal sections were 3.53 ± 0.37 mm, 3.42 ± 0.55 mm, and 3.37 ± 0.54 mm, respectively. In most cases, there was one venous bivalve located in the FV approximately 0-2.71 mm below the junction of the FV and the deep femoral vein. Endothelium covered the luminal and sinusal surfaces of the leaflets. Abundant collagen fibers were found under the endothelial cells beneath the luminal surface of the leaflets. An elastin fiber network was located under the sinus endothelial surface. Smooth muscle cells in the FV extend to the edge of the valve. The venous valve of Macaca fascicularis is similar to that of humans, both morphologically and histologically. However, there is only one venous bivalve and no great saphenous vein in Macaca fascicularis.

El objetivo fue comprender las características anatómicas de la válvula venosa en Macaca fascicularis y compararla con la de los humanos. Fueron disecados bilateralmente los miembros pélvicos (24 miembros de 12 animales) de cadáveres de Macaca fascicularis; las venas femorales (VF) fueron divididas en secciones distal, media y proximal. Se midió el diámetro externo de las VFs en cada sección. Las válvulas venosas se observaron microscópicamente y se tiñeron con H-E y tricrómico. Los datos para describir la válvula venosa humana se obtuvieron desde la literatura. No se encontraron venas safenas magnas entre los 24 miembros inferiores. Los diámetros externos de las VFs en las secciones distal, media y proximal fueron 3,53±0,37 mm, 3,42 mm±0,55, y 3,37±0,54 mm, respectivamente. En la mayoría de los casos, hubo vena bivalva situada aproximadamente 0-2,71 mm debajo de la unión de la VF y la vena femoral profunda. El endotelio cubrió las superficies luminal y sinusal. Se observaron abundantes fibras de colágeno en las células endoteliales bajo la superficie luminal de las válvulas. Una red de fibras de elastina se encontró bajo la superficie del seno endotelial. Las células musculares lisas en las VFs se extiendían hasta el margen de la válvula. La válvula venosa del Macaca fascicularis es similar a la de los seres humanos, morfológica e histológicamente. Sin embargo, sólo hubo una vena bivalvular, y no se observaron venas safenas en Macaca fascicularis.

OBJETIVO: Investigar a possibilidade de terapia multigênica e intervenção por embolização com lipiodol em combinação com quimioterapia focal no tratamento de câncer de fígado VX2 em coelhos. MÉTODOS: Quarenta e cinco coelhos com câncer maior do que 2cm de diâmetro foram distribuídos, aleatoriamente, em cinco grupos (n=9 por grupo). Grupo 1: animais foram tratados com cloreto de sódio 0,9% e no grupo 2 os animais receberam embolização com lipidol. Grupo 3: animais receberam embolização com lipiodol e terapia do gene p53 e grupo 4 animais receberam embolização com lipiodol e terapia do gene TK/CD. Grupo 5: animais receberam embolização com lipiodol e terapia do gene p53 e do gene TK/CD. Ultrassonografia e tomografia computadorizada foram realizadas antes e dez dias após a intervenção terapêutica. RESULTADOS: O modelo VX2 de câncer de fígado foi estabelecido com sucesso em coelhos e a terapia intervencionista foi bem executada. Dez dias após a intervenção terapêutica, uma diferença significativa no volume do tumor foi observada entre os cinco grupos (p<0,05) e diferentes tratamentos poderiam inibir o crescimento do câncer. A inibição do crescimento do cancer foi mais evidente no grupo 5. Análise fatorial revelou que a terapia com gene p53 ou TK/CD e embolia por lipiodol independentemente exerce um efeito inibidor significativo sobre o crescimento do câncer. Além disso, a supressão da taxa de crescimento do tumor foi mais evidente no Grupo 5. CONCLUSÕES: A combinação de terapia gênica com embolização com lipiodol pode inibir efetivamente o crescimento do câncer e prolongar o tempo de sobrevida. Estes resultados demonstram a eficácia da terapia multigênica em combinação com embolização com lipidol no tratamento de câncer hepático.

PURPOSE: To investigate the feasibility of interventional lipiodol embolism and multigene therapy in combination with focal chemotherapy in the treatment of VX2 liver cancer in rabbits. METHODS: Forty five rabbits with cancer larger than 2cm in diameter were randomly divided into five groups (n=9 per group). In Group 1, animals were treated with 0.9% sodium chloride. In Group 2, animals received lipiodol embolism. In Group 3, animals received lipiodol embolism and p53 gene therapy. In Group 4, animals received lipiodol embolism and TK/CD gene therapy. In Group 5, animals received lipiodol embolism and p53 and TK/CD gene therapy. Ultrasonography and CT were performed before and at ten days after interventional therapy. RESULTS: The VX2 model of liver cancer was successfully established in rabbits and interventional therapy smoothly performed. At ten days after interventional therapy, significant difference in the tumor volume was noted among five groups (p<0.05) and different treatments could inhibit the cancer growth. The inhibition of cancer growth was the most evident in the Group 5. Factorial analysis revealed gene therapy with p53 or TK/CD and lipiodol embolism independently exert significantly inhibitory effect on cancer growth. In addition, the suppression on tumor growth rate was the most obvious in the Group 5. CONCLUSIONS: Combination of gene therapy with lipiodol embolism can effectively inhibit the cancer growth and prolong the survival time. These findings demonstrate the effectiveness of multigene therapy in combination with lipiodol embolism in the treatment of liver cancer.