Objective To investigate the relationship between the genetic polymorphisms, which played roles in the pathogenesis of metabolic syndrome (MS), and susceptibility of non-alcoholic fatty liver disease (NAFLD) in Han people in Guangdong province. Methods The subjects were selected from an epidemiologie survey in Guangdong province. Fifty to 117 adult NAFLD patients, who met the criteria of Chinese guideline for diagnosis of NAFLD and had typically clinical, biochemical signs and abdominal ultrasonography, were recruited in the study. By using 1 : 1 matched method of nested case-control study, same numbers of people without NAFLD were included as controls. The genetic analyses was performed by using genomic DNA extracted from peripheral blood. Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) was applied to detect the single nucleotide polymorphisms (SNPs) at 9 sites in 7 candidate genes. Results Most SNPs of the genes were related to the susceptibility of NAFLD. Some of them had positive relation (increasing the risk) such as tumor necrosis factor (TNF)-α-238, adiponectin-45, leptin-2548, peroxisome proliferator-activated receptors (PPAR) γ-161 and phosphatidylethanolamine N-methyltransferase (PEMT)-175. Some had negative relation (decreasing the risk) including adiponectin-276 and hepatic lipase-514. And some had no relation (TNF-α-380 and PPAR g coactivator-1α-482). Conclusion Most cytokines' SNPs of candidate genes discovered in MS patients are related to the susceptibility of NAFLD.

Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.
Animals ; Bombyx ; virology ; Gene Deletion ; Gene Expression Regulation, Viral ; Nucleopolyhedrovirus ; genetics ; physiology ; Transcription, Genetic ; Viral Proteins ; genetics ; metabolism ; Virus Replication

BACKGROUNDThe conventional procedure for screening bioactive components from traditional Chinese medicine is time-consuming, expensive and low efficient. Therefore, some alternative strategies are needed urgently. A novel method for screening anti-platelet aggregation components from oleoresins was developed using chicken thrombocyte extract and high performance liquid chromatography.

METHODSThe anti-platelet aggregation components of oleoresins were combined with receptors, channels and enzymes of chicken thrombocytes under physiological environment. Unbound substances were washed away and bound compounds were eluted using specific phosphate buffered solution (PBS). Compounds released from target sites were collected and analyzed by high performance liquid chromatography and LC-MS. The activity of three compounds which were screened from this model was confirmed using platelet aggregation pharmacology in vivo.

RESULTSThere were four typical compounds that bound to the thrombocytes: 6-gingerol, 8-gingerol, 6-shogaol and 10-gingerol, and all had shown anti-platelet aggregation activities. Eight-gingerol displayed the best anti-platelet aggregation effect.

CONCLUSIONSChicken thrombocyte extract can be used to isolate chemicals that are ligands of the receptor or other bio-targets on the platelet. This may therefore be a simple and efficient method to screen for anti-platelet aggregation compounds from traditional Chinese medicine.

Animals ; Catechols ; isolation & purification ; pharmacology ; Chickens ; Chromatography, High Pressure Liquid ; methods ; Fatty Alcohols ; isolation & purification ; pharmacology ; Ginger ; chemistry ; Medicine, Chinese Traditional ; Plant Extracts ; isolation & purification ; pharmacology ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; isolation & purification ; pharmacology ; Rhizome ; chemistry ; T-Lymphocytes ; metabolism

In order to evaluate the immunogenicity of HPV 58 L1 DNA vaccines, five DNA vaccines had been constructed with pcDNA3.1 vector containing different L1 genes of HPV 58, which were designated as L1h, L1hDeltac, L1S, L1SM and L1wt. The protein expression of DNA vaccines in vitro was tested by Western blot. The ability of forming pseudovirus was evaluated by transfecting DNA vaccine together with pcDNA3.1-h58L2 and pcDNA3.1-GFP into 293FT cells. The neutralizing antibodies and cellular immune response produced in BALB/c mice immunized with the DNA vaccines were detected by using pseudovirus-based neutralization assay and ELISPOT respectively. The results showed that the five DNA vaccines had been successfully constructed; the level of protein expression of L1hDeltac was the highest and those for L1S and L1SM were of medium, while no expressed target protein of L1wt was detected. Only L1S could form the pseudovirus while the other four vaccines could not. L1S and L1h could induce neutralizing antibody. However, the average titer of neutralizing antibody for L1S (1:6,400) was much higher than that for L1h (1:48) and the other three vaccines could not induce neutralizing antibody. No cellular immune response for all five DNA vaccines was detectable by ELISPOT. The results indicated that DNA vaccine against HPV 58 can form pseudovirus in vitro, also can induce high level of neutralizing antibodies. This provides reference for screening HPV vaccine in future.
Animals ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Female ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Models, Genetic ; Neutralization Tests ; Papillomaviridae ; immunology ; Papillomavirus Vaccines ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vaccines, DNA ; genetics ; immunology

Objective To evaluate the preventive and therapeutic effect of oral solution of Niao Du Kang (尿毒康) on acute renal tubular necrosis (ATN) in Sprague-Dawley (SD) rats,and preliminarily approach its mechanisms.Methods The oral solution was composed of traditional Chinese medicinal herbs of Radix et Rhizoma Rhei (大黄),Radix Sanguisorbae Offieinalis (地榆),Radix Salviae Miltiorrhizae (丹参), Radix Astragali seu Hedysari (黄芪),Flos Carthami Tinctorii (红花) and so on.Sixty-four SD male rats were randomly divided into 4 groups:normal control group,ATN model group,verapamil treatment group and traditional Chinese medicine,oral solution of Niao Du Kang treatment group,every group having 16 rats.. The ATN model of SD rats was induced by intramuscular injection of 50% glycerin mixed with isotonic saline solution at the back of bilateral lower extremities.After the model was established in each group ,the levels of blood urea nitrogen (BUN),levels of blood serum creatinine (SCr),renal failure index (RFI) and renal pathological changes were detected at different time points after modeling for 12 and 24 hours,and the therapeutic effect of oral solution of Niao Du Kang was observed.Results After the model establishment, Niao Du Kang oral solution could lower the elevation of blood BUN,SCr,RFI in the model,and compared with the normal control group,there were statistical significances (all P

To find out whether the mutations of HIV-1 Env have influence on the assembly of pseudovirus and their abilities to infect cells, site-directed mutation (A457D)was performed using cycling mutagenesis and selection of mutants with DpnI. Transformation and plasmid purification technologies were used to obtain mutated env clone. Then both the prototype and the mutant were co-transfected with pSG3(delta(env)) to 293FT cells, respectively. Single-cycle infection assay was employed to analyze the effect of the prototype and the mutant on the ability of functional pseudovirus assembly. The transient expression of both the prototype S12-42-1 and mutant S12-42M were confirmed by Western blot essay. The S/CO value was less than 1 for S12-42-1 and 6.65 for S12-42M, demonstrating the functional pseudovirus was generated only for S12-42M. So mutation on HIV-1 Env has influence on the assembly of pseudovirus and their abilities to infect cells.
Amino Acid Sequence ; Base Sequence ; Cell Line ; HIV Infections ; virology ; HIV-1 ; chemistry ; genetics ; physiology ; Humans ; Molecular Sequence Data ; Mutation ; Sequence Alignment ; Virion ; genetics ; physiology ; Virus Assembly ; env Gene Products, Human Immunodeficiency Virus ; chemistry ; genetics ; metabolism

AIM To establish an HPLC-ELSD method for the simultaneous content determination of euscaphic acid,tormentic acid,crategolic acid,corosolic acid,platycodin E,platycodin D3 and platycodin D in Chuanbei Luohan Zhike Granules (Fritillariae cirrhosae Bulbus,Eriobotryae Folium,Platycodonis Radix,etc.).METHODS The analysis of 60％ methanol extract of this drug was performed on a 30 ℃ thermostatic Ultimate XB C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.5％ ammonium acetate flowing at 0.6 mL/min in a gradient elution manner.RESULTS Seven constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.99％-99.78％ with the RSDs of 0.72％-1.72％.CONCLUSION This accurate,simple,specific and reproducible method can be used for the quality control of Chuanbei Luohan Zhike Granules.

In order to investigate the effects of different terms of inhaled nitric oxide (NO) preconditioning with low concentration on the activations of Toll-like receptor 2 and 4 (TLR2/4) in the lung ischemia-reperfusion (IR) injury in mice, we divided the male C57BL mice into five groups: sham (S) group, IR group, NO 1-min preconditioning group (15 ppm NO inhalation for 1 min before ischemia, NO 1-min), NO 10-min preconditioning group (15 ppm NO inhalation for 10 min before ischemia, NO 10-min), NO 60-min preconditioning group (15 ppm NO inhalation for 60 min before ischemia, NO 60-min). The changes of partial pressure of oxygen in artery (PaO2), left lung wet-to-dry weight ratio (W/D), and myeloperoxidase (MPO) in the injured lung were measured in every group at 6th h of reperfusion after 60 min of left lung ischemia. The changes of TLR2/4 activations and plasma TNF-α were measured in this procedure in additional mice. As compared with IR group, PaO2 increased, MPO and W/D decreased evidently after reperfusion in NO 10-min group. The changes in NO 60-min group were similar to those in NO 10-min group. There was no difference between NO 1-min and IR group. In NO inhalation group, the expressions levels of TLR2/4 mRNA and proteins were diminished, TNF-α concentrations were decreased, and the lung injuries were ameliorated effectively. We concluded that short term inhalation of NO protected lung IR injury. But the protective effect of NO was not increased with extension of inhaled NO. Inhaled NO could inhibit the activations of TLR2/4 in the lung after IR injury. TLR signal pathway might contribute to the effect of protection with NO in this model.
Administration, Inhalation ; Animals ; Dose-Response Relationship, Drug ; Lung ; drug effects ; metabolism ; Lung Injury ; metabolism ; prevention & control ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide ; administration & dosage ; Reperfusion Injury ; metabolism ; prevention & control ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Treatment Outcome

OBJECTIVETo study the clinicopathological features and differential diagnoses of mixed epithelial and stromal tumor of the kidney.

METHODSClinical and pathological characteristics of 4 cases of mixed epithelial and stromal tumor of the kidney were studied.

RESULTSThree patients were female and one was male. All patients presented with flank pain and hematuria. Radiologic studies revealed cystic and solid masses involving the kidney. Grossly the tumors had a solid and cyst appearance. Microscopically, the tumors were composed of a mixture of stromal and epithelial elements. The epithelial elements were variable in cell types including cuboidal, hobnail and columnar cells. One case showed Müllerian and intestinal epithelial differentiations. Stromal elements essentially consisted of spindle cells, with thick-walled blood vessels and bands of smooth muscle cells as distinctive features of the tumor. Immunohistochemical staining revealed that the epithelial components were positive for AE1/AE3, whereas the stromal components were positive for ER, PR, and SMA. All patients underwent nephrectomy and were well without evidence of recurrence.

CONCLUSIONSMixed epithelial and stromal tumor of the kidney is a benign neoplasm with distinct histopathological features. It should be distinguished from many other renal neoplasms. Surgical intervention is a preferred therapy.

Actins ; metabolism ; Adult ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Muscle, Smooth ; metabolism ; Neoplasms, Complex and Mixed ; metabolism ; pathology ; surgery ; Neoplasms, Glandular and Epithelial ; metabolism ; pathology ; surgery ; Nephrectomy ; methods ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

Objective: To analyze Internet behaviors of children and adolescents and their influencing factors, and to provide scientific basis for health education guidance and prevention measures. Methods: A stratified cluster sampling method was used. From January to June 2019, 30 955 primary school students in 16 provinces (municipalities), from eastern, central, and western economic zones of China were selected to conduct a questionnaire survey, regarding general demographics, diet and exercise, tobacco, alcohol and sleep, Internet and electronics usage, etc. Results: Among them, 43.1% of students had never been online with girls (44.7%) higher than boys (41.8%)(χ2=21.04, P<0.01); 1.3% of students reported online time for more than 4 hours a day, boys were significantly higher than girls(χ2=15.87, P<0.01); Internet addiction was detected in 2.4% of elementary school students, with boys (3.0%) higher than girls(1.7%)(χ2=331.77, P<0.01), and the trend increases with grade. With sixth grade (3.3%) were significantly higher than children from the first grade (0.8%) and other grades(χ2=163.96, P<0.01). Children’s tendency to Internet addiction was 11.9%, of which boys were 13.0% and girls were 10.6%, boys were significantly higher than girls(χ2=117.66, P<0.01). Pearson correlation analysis revealed that, after controlling gender age, boarding school, family relationship, single parents and regrouped families, smoking, drinking sugary drinks, eating fried foods, and left-behind were positively related to Intenet addiction(r=0.06, 0.07, 0.27, 0.44, 0.07, 0.11, 0.09, P<0.01), while medium and high intensity sports, effective physical education, health education in school, AIDS education and parental restrictions on TV and computer games, were negatively associated with internet addiction(r=-0.74, -0.65, -0.04, -0.05, -0.63, P<0.01). Conclusion The internet addiction of Chinese primary school students is common and increases by year. High-intensity, high-density sports, effective physical education, and health education could help prevent Internet addiction among children.