Danlou Tablet Improves Chronic Intermittent Hypoxia-Induced Dyslipidemia and Arteriosclerosis by HIF-1α-Angptl4 mRNA Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To detect whether Danlou Tablet (DLT) regulates the hypoxia-induced factor (HIF)-1α-angiopoietin-like 4 (Angptl4) mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia (CIH)-induced dyslipidemia and arteriosclerosis.@*METHODS@#The mature adipocytes were obtained from 3T3-L1 cell culturation and allocated into 8 groups including control groups (Groups 1 and 5, 0.1 mL of cell culture grade water); DLT groups (Groups 2 and 6, 0.1 mL of 1,000 µg/mL DLT submicron powder solution); dimethyloxalylglycine (DMOG) groups (Groups 3 and 7, DMOG and 0.1 mL of cell culture grade water); DMOG plus DLT groups (Groups 4 and 8, DMOG and 0.1 mL of 1,000 µg/mL DLT submicron powder solution). Groups 1-4 used mature adipocytes and groups 5-8 used HIF-1 α-siRNA lentivirus-transfected mature adipocytes. After 24-h treatment, real-time polymerase chain reaction and Western blot were employed to determine the mRNA and protein expression levels of HIF-1 α and Angptl4. In animal experiments, the CIH model in ApoE-/- mice was established. Sixteen mice were complete randomly divided into 4 groups including sham group, CIH model group [intermittent hypoxia and normal saline (2 mL/time) gavage once a day]; Angptl4 Ab group [intermittent hypoxia and Angptl4 antibody (30 mg/kg) intraperitoneally injected every week]; DLT group [intermittent hypoxia and DLT (250 mg/kg) once a day], 4 mice in each group. After 4-week treatment, enzyme linked immunosorbent assay was used to detect the expression levels of serum total cholesterol (TC) and triglyceride (TG). Hematoxylin-eosin and CD68 staining were used to observe the morphological properties of arterial plaques.@*RESULTS@#Angptl4 expression was dependent on HIF-1 α, with a reduction in mRNA expression and no response in protein level to DMOG or DLT treatment in relation to siHIF-1 α -transfected cells. DLT inhibited HIF-1 α and Angptl4 mRNA expression (P<0.05 or P<0.01) and reduced HIF-1 α and Angptl4 protein expressions with DMOG in mature adipocytes (all P<0.01), as the effect on HIF-1 α protein also existed in the presence of siHIF-1 α (P<0.01). ApoE-/- mice treated with CIH had increased TG and TC levels (all P<0.01) and atherosclerotic plaque. Angptl4 antibody and DLT both reduce TG and TC levels (all P<0.01), as well as reducing atherosclerotic plaque areas, narrowing arterial wall thickness and alleviating atherosclerotic lesion symptoms to some extent.@*CONCLUSION@#DLT had positive effects in improving dyslipidemia and arteriosclerosis by inhibiting Angptl4 protein level through HIF-1 α-Angptl4 mRNA signaling pathway.

IGF-1R/β-catenin signaling axis is involved in type 2 diabetic osteoporosis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Insulin-like growth factor-1 receptor (IGF-1R) is involved in both glucose and bone metabolism. IGF-1R signaling regulates the canonical Wnt/β-catenin signaling pathway. In this study, we investigated whether the IGF-1R/ β-catenin signaling axis plays a role in the pathogenesis of diabetic osteoporosis (DOP). Serum from patients with or without DOP was collected to measure the IGF-1R level using enzyme-linked immunosorbent assay (ELISA). Rats were given streptozotocin following a four-week high-fat diet induction (DOP group), or received vehicle after the same period of a normal diet (control group). Dual energy X-ray absorption, a biomechanics test, and hematoxylin-eosin (HE) staining were performed to evaluate bone mass, bone strength, and histomorphology, respectively, in vertebrae. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were performed to measure the total and phosphorylation levels of IGF-1R, glycogen synthase kinase-3β (GSK-3β), and β-catenin. The serum IGF-1R level was much higher in patients with DOP than in controls. DOP rats exhibited strikingly reduced bone mass and attenuated compression strength of the vertebrae compared with the control group. HE staining showed that the histomorphology of DOP vertebrae was seriously impaired, which manifested as decreased and thinned trabeculae and increased lipid droplets within trabeculae. PCR analysis demonstrated that IGF-1R mRNA expression was significantly up-regulated, and western blotting detection showed that phosphorylation levels of IGF-1R, GSK-3β, and β-catenin were enhanced in DOP rat vertebrae. Our results suggest that the IGF-1R/β-catenin signaling axis plays a role in the pathogenesis of DOP. This may contribute to development of the underlying therapeutic target for DOP.

Synthesis and cytotoxicity evaluation of 3-amino-2-hydroxypropoxygenistein derivatives / 中国天然药物

Soy isoflavones exhibit various biological activities, such as antioxidant, anti-tumor, anti-inflammatory, and cardiovascular protective effects. The present study was designed to investigate the effects of sixteen synthesized 3-amino-2-hydroxypropoxy genistein derivatives on cell proliferation and activation of Nrf2 (Nuclear factor erythroid 2-related factor 2)/ARE (antioxidant response elements) pathway in human cancer cell lines. Most of the tested compounds exerted greater cytotoxic activity than genistein, as measured by MTT assay. Moreover, compound 8c showed the highest ARE-luciferase reporter activity among the test compounds. It strongly promoted Nrf2 nuclear translocation and up-regulated the expression of total Nrf2 and downstream targets NQO-1 and HO-1 at protein level. The present study may provide a basis for the application of isoflavone derivatives as Nrf2/ARE pathway inducers for cancer therapy and cancer prevention.

Advantages and problems of traditional Chinese medicine in treatment of acute pharyngitis / 中国中药杂志

This paper systematically studies relevant literatures at home and abroad in recent years. China National Knowledge Internet (CNKI) was used to collect the literatures for acute pharyngitis treated with traditional Chinese medicine from January 1, 2006, to December 31, 2016, and the bibliometric method was employed for statistical analysis. A total of 493 papers were preliminarily selected. According to the inclusion criteria and exclusion criteria, 182 eligible articles were selected. According to the evaluation and analysis of the literatures, the Guidelines for Clinical Research of New Drugs is currently used as the common standards for the diagnosis and treatment of acute pharyngitis; Chinese patent medicines are the main traditional Chinese medicine for treating this disease; Decoctions for treatment of this disease include Lonicerae Japonicae Flos, Scutellariae Radix, Platycodonis Radix, Forsythiae Fructus, Glycyrrhizae Radix et Rhizoma, Scrophdlariae Radix, Isatidis Radix, and Ophiopogonis Radix; The bloodletting puncture is the common external therapy. Traditional Chinese medicine and Western medicine have their own characteristics in the treatment of this disease. Western medicine for the treatment of acute pharyngitis are mainly antiviral, antibiotic and glucocorticoid drugs, whose disadvantages are toxicity, side effects, drug resistance and double infections. Traditional Chinese medicine doctors have rich experiences in the treatment of the disease, which is characterized by treatment determination based on syndrome differentiation, safe and reliable medication, significant curative effect, low drug resistance, and wide varieties of traditional Chinese medicine forms, convenient portability and taking, low price, and low toxic and side effects. It is an arduous and significant task to explore traditional Chinese medicine, and study and develop new-type effective drugs.

Synthesis and cytotoxicity evaluation of 3-amino-2-hydroxypropoxyisoflavone derivatives / 中国天然药物

Soy isoflavones exert a wide variety of biological activities, such as antioxidant, anti-inflammatory and anti-cancer properties. Nuclear factor erythroid 2-related factor 2 (Nrf2), a bZip transcription factor, plays a key role in soy isoflavones induced protection against oxidative stress and cancer. To obtain more effective isofavones, a series of 7,4'-bis-(3-amino-2-hydroxypropoxy), 7 or 4'-(3-amino-2-hydroxypropoxy) isoflavone derivatives have been synthesized as potential antitumor agents and Nrf2/ARE (antioxidant response element) activators. The cytotoxicity of these compounds in human cancer cell lines MDA-MB-231, HT-29, HCT116, HepG2 and 7402 was tested by MTT assay. In this study, the cytotoxicity of compound 3b exhibited highest cytotoxic activity and at the safety dose range, it also strongly up-regulated antioxidant response element (ARE)-luciferase reporter activity. In addition, compound 3b induced Nrf2 nuclear translocation and upregulated its downstream target genes NQO-1 and HO-1 at protein level. Taken together, our results suggest that compound 3b could be a potential agent for cancer themotherapy or cancer chemoprevention.

Clinical significance of serum carbohydrate antigen 125 in acute exacerbation of chronic obstructive pulmonary disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the serum level of carbohydrate antigen 125 (CA125) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and its relation with pulmonary hypertension.</p><p><b>METHODS</b>Forty-six patients with AECOPD complicated by pulmonary hypertension, 46 with AECOPD and 38 healthy control subjects were examined for their clinical data, pulmonary function, echocardiographic findings, and serum levels of lung tumor markers and brain natriuretic peptide (BNP).</p><p><b>RESULTS</b>Compared with the healthy control group, COPD patients with or without pulmonary hypertension showed significantly decreased pulmonary function (P<0.05), especially in those with AECOPD and concurrent pulmonary hypertension (P<0.05). Serum CA125 level was obviously higher in AECOPD group than in the healthy control group, and further increased in AECOPD patients with pulmonary hypertension (P<0.05). The levels of lung tumor markers (CEA, NSE, CYFRA and PROGRP) were similar among the 3 groups (P>0.05). The serum level of BNP in patients with AECOPD and concurrent pulmonary hypertension was significantly higher than that in patients with AECOPD (P<0.05). Pearson linear correlation analysis showed that serum CA125 was positively correlated with pulmonary artery systolic pressure and BNP in AECOPD patients with pulmonary hypertension (P<0.01).</p><p><b>CONCLUSION</b>Serum CA125 may serve as a serological index to identify AECOPD patients with pulmonary hypertension.</p>

Molecular epidemiological analysis of ECHO7 virus isolated from sewage water in Yunnan Province, China / 病毒学报

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.

Study on the molecular typing and epidemiology of non-polio enterovirus in Yunnan Province, China during 2006-2010 / 病毒学报

In order to explore the genotype distribution and molecular evolution of non-polio enterovirus (NPEVs)in Yunnan Province,the People's Republic of China, we sequenced and analyzed the partial VP1 coding region of 105 NPEVs isolated from acute flaccid paralysis (AFP) surveillance in Yunnan province during a 5- year study period from 2006 to 2010. The viral genomes of 105 NPEVs were translated to corresponding amino acid sequences and compared with those of the prototype strains, and the phylogenetic tree was constructed among these VP1 nucleotide sequences and other prototype strains from GenBank. Analysis showed that 18 isolates were classified into 7 serotypes of human enterovirus A species, while 77 isolates into 22 serotypes of B and 10 isolates into 4 serotypes of C species. However, we did not isolate any viruses which belonged to human enterovirus D species. Thus, under AFP surveillance, human enterovirus B species accounted for 73. 3% of the 105 isolates and was considered as the predominant one,followed by human enterovirus A(17. 1%) and human enterovirus C(9. 5%). Phylogenetic analysis showed that various serotypes of the virus and the corresponding prototype strains or other representative strains clustered into the same grooup, however, Yunnan strains and prototype strains were located in the different branches (except CA2,EV90 and EV76). The degree of variation was different even among the same genotype strains. This report showed that different genotype strains spread widely in Yunnan Province.

Investigation on one vaccine-derived poliovirus (VDPV) case in Yunnan Province / 病毒学报

To find out epidemiological feature of paralytic cases caused by type 2 vaccine-derived poliovirus (VDPV) and the excretion status of the case and to explore the enterovirus infection status among healthy children under five years old around the case in Zhaotong city, Yunnan Province in 2010. Field epidemiological studies at the epidemic area were conducted and a total of 108 stool samples were collected, three from the case, seven from the close contacts and 98 from the healthy children. VDPV was not isolated again from the case; Sabin-like PV strains or VDPV were not isolated from the close contacts and the healthy children; An active search for acute flaccid paralysis cases was conducted in the area, which indicated that the VDPV did not cause virus circulation in local area. Twenty one (20.0%) NPEVs were isolated from 105 stool samples. Among the 21 NPEV isolates, 11 isolates (52.4%) were HEV-A (3 serotypes), 10 isolates (47.6%) were HEV-B (4 serotypes).

Molecular typing of enteroviruses from healthy children in the border areas of Yunnan Province and Myanmar and the genetic characteristics of ECHO7 and ECHO13 in 2009 / 病毒学报

To explore the enteroviruses surveillance among healthy children under 15 years old in the border areas of Yunnan Province and Myanmar in 2009. The stool samples were collected from the healthy children under 15 years old who came from the border areas of Myanmar and Yunnan Province, virus isolation and sequencing were conducted for all the 271 samples. 6 strains of polioviruses (PVs) were detected from 271 stools with an isolation rate of 2.8%, which belonged to vaccine strains and 24 non-polioviruses (NPVs) were detected with an isolation rate of 8.9%. 24 NPVs belonged to human enterovirus group B (HEV-B) with 6 serotypes, HEV-A, HEV-C and HEV-D viruses were not isolated. Among them, 13 NPVs were E7 (54.17%) and 5 NPVs were E13 (20.83%). Our results showed that the enterovirus carrying rate in the border areas of Yunnan province was higher than the rate of routine acute flaccid paralysis (AFP) detection system. The HEV-B viruses were the only enteroviruses isolated. The phylogenetical analysis showed that Echovirus 7(E7) and 13 (E13) exhibited genetic polymorphism.

Status of enterovirus infection and molecular identification among healthy children at the areas bordering Myanmar, in Yunnan province,China / 中华流行病学杂志

Objective To explore the enterovirus infection status among healthy children under 15 years old in the border areas of Yunnan province that connecting Myanmar.Methods A total of 319 stool samples were collected from healthy children in the 10 entrance ports.Enterovirus was isolated from these stool samples and then poliovirus and adenovirus were serotyped by neutralization test using specific anti-sera.All the non-polio enteroviruses(NPEVs)were identified by partial sequencing of VP1 gene.Results All 53 enterovirus were isolated from 319 stool samples and 16.6% of them carried the virus.23 polio virus(PVs)and 30 NPEVs were isolated with rates of carrying the virus were 7.2% and 9.4% respectively.4 adenovirus were also isolated with a rate as 1.25%.1 isolate could not be amplified by any Pan-enterovirus primers or by RT-PCR so was not able to be sequenced.The results of NPEVs sequencing showed that:1 isolate(3.3%)was classified into 1 serotype of HEV-A while 20 isolates(66.7%)were classified into 11 serotypes of HEV-B and 8 isolates(26.7%)were classified into 3 serotypes of HEV-C.However,we could not isolate any viruses that belong to HEV-D.nt.Result from the aa identify calculation showed that the nt and aa identification between isolates and corresponding standard strains were more than 75% and 85% respectively.The findings were similar to the international standards.Conclusion Our results showed that the rate of carrying the enterovirus especially poliovirus in some areas of Yunnan province that bordering Myanmar was higher than that of rate through the routine acute flaccid paralysis detection system.Of the enterovirus isolated,HEV-B group appeared the predominant with the wide spread of enterovirus serotype.Some newer enterovirus were also detected such as EV73(2 strains),EV75(1 strain),EV80(1 strain)and EV96(4 strains).

Molecular identification of human enterovirus 73 in Yunnan Province, the People's Republic of China / 病毒学报

Molecular typing was conducted according to the reported methods for 2 enteroviruses that were isolated from healthy children in the border areas of Yunnan Province with Myanmar. RT-PCR and sequencing were performed with 292/222 primers according to the Oberste's methods. The resulting sequences were blasted against the Genbank database and compared with all available enterovirus database. Analysis of homology at nucleotide and amino acid level identically suggested that the two enteroviruses are human enterovirus 73.