Construction and preliminary application of drug carrier system based on yeast membrane vesicles / 药学学报

As an edible eukaryotic microorganism, Saccharomyces cerevisiae has the characteristics of high safety, rapid proliferation, low cost, easy transformation, etc. It has been widely used to produce vaccines, antibodies, insulin, etc. Up to now, yeast components, such as cell wall and yeast microcapsules, have been widely used in the treatment of tumors, inflammatory virus infection, post-traumatic osteoarthritis and other diseases. Among them, the components of yeast cell membrane are relatively simple and stable, which are easy to be extracted on a large scale. Therefore, yeast cell membrane material was used to construct yeast membrane vesicle nanosystem, and its biomedical application was preliminarily explored. In this study, Saccharomyces cerevisiae membrane vesicle (SMV) was prepared by co-extrusion method, and the particle size and surface potential of SMV, drug loading and release characteristics, stability, cell safety, and in vitro therapeutic effect were investigated. The results showed that the average particle size of SMV was 185.1 nm. Curcumin and silica nanoparticles were effectively encapsulated by co-incubation and ultrasonic methods, and the characteristics of cell membrane proteins were maintained. Moreover, SMV had good stability and biocompatibility. In addition, SMV could be effectively uptaken by macrophages RAW 264.7, and curcumin loaded SMV could effectively eliminate reactive oxygen species (ROS). In conclusion, the yeast plasma membrane vesicles prepared in this study could effectively deliver curcumin drugs and encapsulate nanoparticles, and could be effectively absorbed by macrophages and effectively eliminate ROS, providing new ideas and new methods for biomedical applications of yeast membrane materials.

MicroRNA-770-5p contributes to podocyte injury via targeting E2F3 in diabetic nephropathy

Diabetic nephropathy (DN) has been identified as the major cause of end-stage renal disease (ESRD) in most developed countries. MicroRNA-770-5p depletion could repress high glucose (HG)-triggered apoptosis in podocytes, and downregulation of E2F transcription factor 3 (E2F3) could facilitate podocyte injury. Nevertheless, whether E2F3 is involved in miR-770-5p knockdown-mediated improvement of DN is still unclear. The expression levels of miR-770-5p and E2F3 were detected in HG-treated podocytes by RT-qPCR. The expression levels of E2F3, apoptosis-related proteins Bcl-2 related X protein (Bax), B-cell lymphoma-2 (Bcl-2), Bad, apoptotic peptidase activating factor 1 (APAF1), C-caspase3, C-caspase7, and C-caspase9 were detected by western blot assay. The effects of miR-770-5p and E2F3 on HG-treated podocytes proliferation and apoptosis were detected by CCK-8 and flow cytometry assays. The interaction between miR-770-5p and E2F3 was predicted by Targetscan, and then verified by the dual-luciferase reporter assay. MiR-770-5p was upregulated and E2F3 was downregulated in HG-treated podocytes. MiR-770-5p inhibited proliferation and promoted apoptosis and E2F3 promoted proliferation and suppressed apoptosis in HG-treated podocytes. E2F3 is a target gene of miR-770-5p and it partially abolished the effect of miR-770-5p in HG-triggered proliferation and apoptosis of podocytes. MiR-770-5p deficiency blocked HG-induced APAF1/caspase9 pathway via targeting E2F3 in podocytes. We firstly confirmed that E2F3 was a target of miR-770-5p in podocytes. These findings suggested that miR-770-5p expedited podocyte injury by targeting E2F3, and the miR-770-5p/E2F3 axis might represent a pathological mechanism of DN progression.

Effect of Serum Containing Buyang Huanwu Tang on Endothelial-to- mesenchymal Transition of Human Pulmonary Artery Endothelial Cells / 中国实验方剂学杂志

Objective:To investigate the regulatory effect of serum containing Buyang Huanwu Tang on endothelial-to-mesenchymal transition(EndMT) in human pulmonary artery endothelial cells (HPAEC), and make further analysis on its mechanism from the perspective of the signal transduction of Jagged1/Notch1. Method:Rabbit serum containing Buyang Huanwu Tang was prepared by gavage with dosage of 53.36 g·kg-1·d-1, and blank serum was prepared by gavage with same volume of normal saline. The HPAECs cultured in vitro, EndMT model was established by the transforming growth factor-β1(TGF-β1) induced, which were divided into five groups:the control group (10%blank serum), the model group (10%blank serum+TGF-β1), the serum containing high-dose Buyang Huanwu Tang group (10%medicated serum + TGF-β1), the medium-dose group (5%medicated serum + 5%blank serum medicated + TGF-β1) and the low-dose group(2.5%medicated serum+7.5%blank serum+ TGF-β1). Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) method. Cell migration was detected by transwell and scratch assay. The endothelial markers platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31), vascular endothelial cadherin (VE-cadherin) and the mesenchymal markers fibroblast-specific protein 1 (FSP1), α-smooth muscle actin (α-SMA) were observed by immunofluorescence assay. The expression levels of Notch1, Jagged1 and CBF1 were detected by Western blot assay. Result:Compared with the control group, the proliferation and migration abilities of the HPAEC cells in model group were enhanced (Pα-SMA were increased. Further study found that the expressions of Notch1, Jagged1 and CBF1 were up-regulated (PPPα-SMA were on the decline. The expressions of Notch1, Jagged1 and CBF1 were also significantly lower than those in model group (PPConclusion:The serum containing Buyang Huanwu Tang can partly inhibit the EndMT in human pulmonary artery endothelial cells, which may be related to the regulation effect of Jagged1/Notch1 signaling.

Dietary behavior problems of 2-7 yearold autistic children in Urumqi / 中国学校卫生

Objective@#To compare the differences of dietary behavior between autistic children and normal children, and to explore the dietary behavior problems and their influencing factors in autistic children, so as to provide theoretical basis for improving dietary behavior problems and guiding scientific feeding of autistic children.@*Methods@#113 autistic children aged 2-7 from a hospital and an autism rehabilitation training institution in Urumqi were selected as the case group and 223 children aged 2-7 from a hospital in Urumqi as the control group.@*Results@#The results showed that there were significant differences between the two groups in the main type of care, caregiver education, caregiver occupation and monthly per capita economic income(P<0.05). The scores of picking food, food response, unhealthy eating habits, overeating response and emotional eating were significantly higher in the case group than in the control group, while the scores of extrinsic eating and active eating ability were significantly lower in the case group than in the control group(P<0.05).@*Conclusion@#The main dietary problems of autistic children include severe picky eating, food response to different degrees, over fullness response, bad eating behavior, emotional eating and so on.

Long-term outcomes following mitral valvuloplasty versus replacement for native valve endocarditis: A case control study / 中国胸心血管外科临床杂志

@#Objective    To compare long-term outcomes following mitral valvuloplasty (MVP) and mitral valve replacement (MVR) for native valve endocarditis (NVE). Methods    Between November 1993 and August 2016, consecutive 101 patients with NVE underwent mitral surgery in our department, MVP for 52 patients and MVR for 49 patients. There were 69 males and 32 females at age of 38.1±14.9 years. The mean follow-up was 99.4±75.8 months. Results    There was no statistical difference in cardiopulmonary bypass time, aortic cross-clamp time, in-hospital mortality, duration of mechanical ventilation, ICU stay or hospital stay after surgery between the two groups. Survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 97.6%, 97.6%, 97.6% for MVP, and 93.5%, 84.3%, 84.3%, 66.2% for MVR with a statistical difference between the two groups (P=0.018). There was no stroke in the patients with MVP during follow-up periods. However, stroke-free survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 93.9%, 89.4%, 70.2% for MVR patients with a statistical difference between the two groups (P=0.023). There was no statistical difference in recurrence of infection, perivalvular leakage and reoperation between the two groups. Composite endpoint-free survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 97.6%, 92.9%, 92.9% for MVP, and 91.3%, 79.6%, 75.8%, 51.0% for MVR with a statistical difference (P=0.006). Conclusion    MVP is associated with better outcomes than MVR in the patients  with NVE; generalizing MVP technique in the patients with NVE is needed.

Potential Value ofAgglutinin-recognized Glycopatterns in Urinary Protein on Differential Diagnosis of Diabetic Nephropathy and Nondiabetic Renal Disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Diabetic nephropathy (DN) is the most common and serious microvascular complication of diabetes. To date, the gold standard for identifying DN and nondiabetic renal disease (NDRD) is a renal biopsy; however, there is currently no reliable diagnostic marker to identify DN and NDRD in a noninvasive manner. This study aimed to investigate the different glycopatterns in urine specimens of DN patients and NDRD patients for a differential diagnosis.</p><p><b>METHODS</b>In total, 19 DN patients and 18 NDRD patients who underwent renal biopsies between March 2015 and March 2016 at the Chinese People's Liberation Army General Hospital were enrolled in this study. A lectin microarray was used to investigate the glycopatterns in the urinary protein of the 37 patients. Ratio analysis and one-way analysis of variance were used to screen altered glycopatterns. Then, the altered glycopatterns between the DN and NDRD groups were verified by a urinary protein microarray among another 32 patients (15 with DN and 17 with NDRD), and receiver operating characteristic (ROC) curve analysis was used to determine the diagnostic value of the altered glycopatterns in differentiating DN and NDRD. Finally, lectin blotting was used to evaluate the altered glycosylation in protein level.</p><p><b>RESULTS</b>The result of lectin microarrays revealed that the relative abundance of the (β-1,4)-linked N-acetyl-D-glucosamine (GlcNAc) recognized by lectin Datura stramonium agglutinin (DSA) was significantly higher in urinary protein in DN patients than that in NDRD patients (fold change >1.50, P < 0.001). Subsequently, the results of urinary protein microarrays were consistent with lectin microarrays (P < 0.05). Furthermore, the ROC curve showed that glycopatterns could effectively distinguish DN from NDRD patients (area under the ROC curve = 0.94, P < 0.001). DSA lectin blotting showed that glycoproteins, with a molecular weight of approximately 50,000, demonstrated a difference in urine samples between DN patients and NDRD patients.</p><p><b>CONCLUSIONS</b>The relative abundance of (β-1,4)-linked GlcNAc recognized by lectin DSA and urinary glycoprotein with a molecular weight of approximately 50,000 are significantly different between DN and NDRD patients, indicating that the glycopatterns could be used as potential biomarkers for a differential diagnosis.</p>

Comparison of nerve combing and percutaneous radiofrequency thermocoagulation in the treatment for idiopathic trigeminal neuralgia / Comparação entre neurólise interna e termocoagulação percutânea por radiofrequência no tratamento da neuralgia idiopática do trigêmeo

ABSTRACT INTRODUCTION: Idiopathic trigeminal neuralgia (ITN) is a common pain disease in elderly people. Many methods have been used to alleviate the pain of patients, but few studies in the literature have compared the effect of nerve combing and percutaneous radiofrequency thermocoagulation. OBJECTIVE: The purpose of this study was to describe and evaluate the clinical outcome of idiopathic trigeminal neuralgia after nerve combing (NC) and compare them with those obtained using percutaneous radiofrequency thermocoagulation (RF). METHODS: The study included 105 idiopathic trigeminal neuralgia patients with similar symptom, age and underlying disease, which were divided into two groups. One group was treated by nerve combing (50 patients), the other by RF (55 cases). All patients were considered medical failures prior to the surgeries. A questionnaire was used to assess the long-term outcomes: pain relief, recurrence, complication and need for additional treatment. RESULTS: The median duration of follow-up in both groups was 90 months. Satisfactory relief was noted in 41 patients (82%), 5 patients (10%) initially experienced pain relief, then recurred, and four patients (8%) were designated poor among the group NC. In the group RF, satisfactory relief was noted in 42 patients (76.4%). There were eight "pain free with recurrence patients (14.5%) and 5 poor cases (9.1%). No statistically significant differences existed in the outcomes between both groups (p > 0.05). Postoperative morbidity included dysesthesia, diplopia, partial facial nerve palsy, hearing loss, tinnitus, cerebrospinal fluid leak, meningitis and mortality. CONCLUSION: Nerve combing and RF are both satisfactory treatment strategies for patients with ITN. Because of the higher risk of sensory morbidity and surgical risk as open surgery, RF is preferred as the recommended procedure for patients with ITN.

Resumo Introdução: A neuralgia idiopática do trigêmeo (NIT) é uma condição dolorosa comum em idosos. Muitos métodos têm sido usados para aliviar a dor dos pacientes, mas poucos estudos na literatura compararam o efeito de neurólise interna e termocoagulação percutânea por radiofrequência. Objetivo: O objetivo desse estudo foi descrever e avaliar o desfecho clínico de pacientes com neuralgia idiopática do trigêmeo após neurólise interna (NI) e compará-los com os obtidos usando termocoagulação percutânea por radiofrequência (RF). Método: O estudo incluiu 105 pacientes com NIT com sintomas, idade e doenças de base semelhantes, que foram divididos em dois grupos. Um grupo foi tratado por neurólise interna (50 pacientes) e o outro por RF (55 casos). Todos os pacientes haviam sido considerados fracassos terapêuticos antes das cirurgias. Um questionário foi utilizado para avaliar os resultados a longo prazo: alívio da dor, recorrência, complicações e necessidade de tratamento adicional. Resultados: A duração média do acompanhamento foi de 90 meses em ambos os grupos. Alívio satisfatório foi observado em 41 pacientes (82%); cinco pacientes (10%) experimentaram alívio inicial da dor, porém seguido de recrudescimento, e quatro pacientes (8%) apresentaram desfecho desfavorável no grupo NI. No grupo de RF, alívio satisfatório foi observado em 42 pacientes (76,4%). Houve oito pacientes livres de dor, com recorrência ''LDR'' (14,5%) e cinco casos com desfecho desfavorável (9,1%). Não houve diferenças significantes nos resultados entre os dois grupos (p > 0,05). Morbidade pós-operatória incluiu disestesia, diplopia, paralisia parcial do nervo facial, perda auditiva, tinnitus, fístula liquórica, meningite e óbito. Conclusão: Neurólise interna e RF são estratégias satisfatórias de tratamento para os pacientes com NIT. Em decorrência da maior morbidade sensorial e maior risco cirúrgico em uma cirurgia aberta, a RF é o procedimento mais indicado para pacientes com NIT.

Expression and significance of E-cadherin, N-cadherin, transforming growth factor-β1 and Twist in prostate cancer

OBJECTIVE@#To study the expression of E-cadherin, N-cadherin, TGF-β1 and Twist protein and investigate its significance in the occurrence and development of prostate cancer.@*METHODS@#The expression of E-cadherin, N-cadherin, TGF-β1 and Twist protein in 59 prostate cancer tissues and 21 adjacent tissues were detected by immunohistochemical SABC staining, and the correlation with clinicopathological features was analyzed.@*RESULTS@#Positive rates of E-cadherin, N-cadherin, TGF-β1 and Twist were 32.2%, 54.2%, 71.2% and 74.6%, respectively, in prostate cancer tissues and 85.7%, 9.52%, 19.0% and 9.52%, respectively, in cancer-adjacent tissues, with significant differences between the two groups (P20μg/L group, but the positive expression of Twist was not significant between groups. The expression of E-cadherin was highly negatively correlated with that of N-cadherin and also highly negatively correlated with that of Twist. The expression of TGF-β1 was correlated with those of E-cadherin, N-cadherin and Twist.@*CONCLUSIONS@#The reduced expression of E-cadherin, abnormal expression of N-cadherin, transformation form E-cadherin to N-cadherin and the increased expression of TGF-β1 and Twist play an important role in the occurrence and development of prostate cancer.

Primary Study of Producing UMP by Overexpressing URA 5 and URA 3 Genes in Saccharomyces cerevisiae / 中国生物工程杂志

To increase the biotransfomation efficiency from the orotic acid to the uridine 5'-monophosphate(UMP),URA5 gene encoding orotate phosphoribosytransferase was amplified from Saccharomyces cerevisiae BY4742 by PCR,then it was inserted into the expression vector pYX212(contained orotidine monophosphate decarboxylase gene URA3)and the pYX212-URA5 was transformed into Saccharomyces cerevisiae BJX12 by electroporation.The recombinant strain was elementarily used to convert orotic acid to UMP.The results showed that pYX212-URA5/BJX12 could accumulate 7mmol/L UMP from 32mmol/L orotic acid in 26h,significantly higher than both control groups pYX212/BJX12(2.7mmol/L) and BJX12(2.4 mmol/L).

Effects of sucrose and light on the growth and production of secondary metabolites in Pueraria phaseoloides hairy roots / 生物工程学报

Effects of sucrose concentrations and light on the growth and production of total isoflavones and puerarin in Pueraria phaseoloides hairy roots cultured onto solid MS media supplemented with 1%, 3%, 5%, 7% and 9% sucrose, respectively, were investigated. The results showed that among the sucrose concentrations tested, 3% sucrose in the medium enhanced the growth and stimulated accumulation of total isoflavones and puerarin in P. phaseoloides hairy roots, After cultured for 20 days, the biomass of hairy roots reached 0.48 g (DW dry weight)/flask and its contents of total isoflavones and puerarin were 25.44 mg/g (DW) and 11.64 mg/g (DW), respectively. In comparison with 3% sucrose, the dry weight proliferation of hairy roots cultured with 5% sucrose was increased by 7.0%, while cultured with 1%, 7% and 9% sucrose, the dry weight proliferation of hairy roots was decreased by 62.4%, 42.8% and 65.3%, their total isoflavones content was decreased by 57.4%, 13% and 33.4% and their puerarin content was decreased by 47.9%, 15.8% and 35.1%; but their content of total soluble sugars was increased 0.52, 1.45 and 1.54 times, respectively. Compared with hairy roots in blue light and white light, the biomass of hairy roots cultured in the dark for 30 days was 0.83 g (DW)/flask and was increased by 37.1% and 23.3%, respectively. The content of total isoflavones in hairy roots cultured in white light was as much as 1.15 times and 1.19 times that in blue light and in the dark, respectively. It was also observed that hairy roots cultured in blue light and white light partly became light green and that blue light could inhibit accumulation of puerarin in hairy roots and the puerarin content in hairy root cultured in white light and in the dark were 1.61 times and 1.52 times that in blue light, respectively.