"Omics" and bioinformatics have brought new ideas to the study of traditional Chinese medicine. This study used metabonomics and network pharmacology to investigate the pharmacodynamic basis and regulation of Qishen Yiqi dropping pill (QDP) improving cardiac energy metabolism in rats with heart failure (HF). ¹H NMR metabonomics analysis showed that eight metabolites, including carnitine, glutamine, creatine, proline, homocitrulline, lactic acid, taurine and alanine appeared significant callback after QDP treatment for HF. The results indicate that QDP regulates the metabolism of carbohydrate, lipid, ATP and protein. The animal experiment was conducted in accordance with the regulations of the Ethics Committee for Experimental Animal Management and Animal Welfare of Institute of Materia Medica, Chinese Academy of Medical Sciences. A "drug-component-target-disease" network was established using network pharmacology, and the "component-target" sub-network related to the above energy metabolism processes was extracted by combining metabonomics results. Results revealed 79 chemical compounds and 47 potential targets of QDP involved in the regulation of energy metabolism, and identified key chemical components including ursolic acid, notoginsenoside G, ginsenoside-Rh1, and core targets such as INS, PPARG, and AKT1. The results also demonstrated the complex multi-target and multi-component relationship between QDP and HF from the perspective of energy metabolism. The molecular docking technique verified a strong interaction between some targets and chemical compounds, with affinities less than -5 kcal·mol^-1. The results of this study provide useful information for the clinical application, development, and utilization of QDP.

Objective: To analyze the clinical effect of traditionaldog-days acupoint application and Magic Acupuncture Patch (Manji) in the prevention and treatment of chronic bronchitis (CB) in remission stage in the past five years, and explore the principle of action and effective stimulation, to provide the evidence for treating CB by acupoint application. Methods: A retrospective study was conducted on 405 patients with CB who met the inclusion criteria. All patients were treated with dog-days acupoint application or Magic Acupuncture Patch between 2013 and 2017. The clinical data of 405 patients were statistically analyzed to compare the prevention and treatment effects of dog-days acupoint application and Magic Acupuncture Patch, and different degrees of stimulation of dog-days acupoint application. Results: Among the dog-days acupoint application groups, the total effective rate was 63.6％ in the light stimulation group, 93.1％ in the moderate stimulation group, and 94.8％ in the strong stimulation group. The differences in the total effective rate between the light stimulation group and the moderate stimulation group, as well as the strong stimulation group, were statistically significant (both P<0.05). There was no significant difference in the total effective rate between the moderate stimulation group and the strong stimulation group (P>0.05). The total effective rate was 83.9％ in the dog-days acupoint application group, versus 45.4％ in the Magic Acupuncture Patch group, and there was a statistically significant difference between the two groups (P<0.05). Conclusion: The efficacy of dog-days acupoint application in the prevention and treatment of CB is better than that of Magic Acupuncture Patch; the degree of stimulation is the basis for the effect of dog-days acupoint application on prevention and treatment of CB, and the moderate and strong stimulations are more appropriate.

Studies have found that metformin is not only the preferred drug for lowering blood sugar, but also shows lipid-lowering and weight-loss effects. The purpose of this study was to use a hyperlipidemia hamster model to investigate the lipid-lowering effect of metformin and its effect on important metabolic pathways in lipid metabolism disorders. Fifty golden hamsters were divided into a control group, a model group, metformin high- and low-dose groups, and a simvastatin group. A high-fat diet was fed for 1 week to create the model, and then drug was administered for 11 weeks with the high-fat diet. Serum was taken for measurement of blood lipid and blood glucose at 2, 6, and 9 weeks after administration, and at weeks 3, 5, and 9 feces and urine were collected for ¹H NMR metabolomics tests. After 11 weeks of intravenous injection of [U-¹³C₆] glucose, serum was collected for a ¹³C NMR metabolic flux test. The results showed that the administration of metformin can significantly reduce blood lipids and glucose levels and can significantly affect metabolic pathways such as sugar metabolism, lipid metabolism, ketone metabolism, amino acid metabolism, and intestinal flora metabolism. The results of the metabolic flux analysis showed that the high-fat diet reduced the metabolism of tricarboxylic acids by 37.48%. After administration of low and high doses of metformin the metabolism of tricarboxylic acid increased by 98.14% and 143.10%, respectively. After administration of simvastatin tricarboxylic acid metabolism increased by 33.18%. The results indicate that metformin has a significant effect on promoting energy metabolism. This study used a combination of metabolomics and metabolic flow to explore the effect of metformin on lipid metabolism disorders and quantifies changes in the key pathway of energy metabolism-the tricarboxylic acid cycle. This study provides useful information for the study of the efficacy and mechanism of metformin, as well as a practical technical method for the screening of lipid-lowering drugs based on a hamster model.

In genetic modification of plants, once the transformants are obtained, selection markers are no longer required in mature plants. At present, the Cre/lox site-specific recombination system is most widely used to eliminate the selectable marker genes from the transgenic plants. In this study, attempt was made to favour the selection of marker-free plants in the re-transformation method. Green fluorescent protein (GFP) can be directly visualized in living cells, tissues or organisms under UV illumination. This advantage of GFP is exploited in the development of a practical approach in which GFP is used as a visual marker to monitor the removal of the selectable marker gene from transgenic plants. For that purpose, the pGNG binary vector was constructed, in which the GFP gene (gfp) was linked to the expression cassette Nos P-nptII-NosT and the two units were cloned between two directly-orientated lox sites. The CaMV 35S promoter was placed before the first lox site and used to drive GFP expression. The beta-glucuronidase gene (gus) of Escherichia coli was cloned behind the second lox site without a promoter, thus would not be expressed in this position. Tobacco plants were first transformed with pGNG and selected on kanamycin (Kan)-containing media. Regenerated transgenic shoots were readily singled out by GFP fluorescence. The GFP-expressing plants were then re-transformed with pCambia1300-Cre containing hygromycin phosphotransferase gene (hpt) as a selectable marker gene. The Cre-mediated recombination resulted in the elimination of lox-flanked genes, herein gfp and nptII, from the plant genome and brought the GUS gene next to the 35S promoter. Our data demonstrated that transgenic plants free of nptII were easily selected by monitoring the loss of green fluorescence, and at the same time, GUS (here as a target protein) was expressed in the nptII-free plants. Finally, hpt and cre were removed from the progenies of the nptII-free plants by gene segregation.
Genetic Markers ; Green Fluorescent Proteins ; genetics ; Plants, Genetically Modified ; genetics ; Plasmids ; Recombination, Genetic ; Tobacco ; genetics

AIMTo investigate the protective effects of shark hepatic stimulator substance (sHSS) against acute hepatic injury induced by acetaminophen (AAP) in mice.

METHODSAcute hepatic injury model of Balb/c mice was induced by a single intraperitoneal injection of AAP (200 mg.kg-1, i.p.). Serum ALT and AST activities were analyzed. The changes of microstructure and ultrastructure of hepatocyte were observed under optical and electronic microscope. The hepatocyte apoptosis was analyzed by flow cytometer and the expression level of Fas mRNA was determined by RT-PCR.

RESULTSThe activities of serum ALT and AST were significantly decreased and both necrosis and inflammatory infiltration were improved in the mice treated with sHSS 3.0 and 1.5 mg.kg-1. sHSS (3.0 mg.kg-1) prevented the ultrastructural changes of hepatocytes caused by AAP, decreased the percentage of apoptotic cells, and downregulated the expression level of Fas mRNA.

CONCLUSIONsHSS protected hepatocytes from AAP-induced injury, which might be associated with its protection of the mitochondria and inhibition of apoptosis and expression of Fas mRNA in hepatocytes.

Acetaminophen ; Animals ; Apoptosis ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; pathology ; Female ; Growth Substances ; isolation & purification ; pharmacology ; Mice ; Mice, Inbred BALB C ; Peptides ; isolation & purification ; pharmacology ; Protective Agents ; pharmacology ; RNA, Messenger ; genetics ; Random Allocation ; Sharks ; fas Receptor ; biosynthesis ; genetics

OBJECTIVETo investigate the effect of transurethral resection of the prostate (TURP) in the treatment of advanced prostate cancer with bladder outlet obstruction (BOO).

METHODSWe included in this study 43 cases of advanced prostate cancer with BOO treated by TURP, and analyzed their IPSS, maximum urinary flow rate and relevant risk factors pre-operatively and at 3 and 12 months after TURP.

RESULTSCompared with the baseline, IPSS and the maximum urinary flow rate of the patients showed significant differences 3 months after surgery ([19.60 +/- 0.41] score vs. [9.58 +/- 0.33] score, [4.93 +/- 0.68] ml/s vs. [8.96 +/- 0.47] ml/s, P < 0.05), but not at 12 months ([15.73 +/- 0.66] score, [5.67 +/- 0.44] ml/s). In multiple regression analysis, a good outcome was associated with pre-operative acute urinary retention, while poor prognosis with hormone-refractory prostate cancer.

CONCLUSIONIn the treatment of advanced hormone-refractory prostate cancer with BOO, TURP can reduce IPSS and increase the maximum urinary flow rate in the early period after surgery, but its long-term effect is not so desirable. Meanwhile the operation itself may bring about relevant complications and reduce the patient's quality of life.

Aged ; Aged, 80 and over ; Feasibility Studies ; Humans ; Male ; Middle Aged ; Prostatic Neoplasms ; surgery ; Transurethral Resection of Prostate ; Treatment Outcome ; Urinary Bladder Neck Obstruction ; surgery

OBJECTIVETo investigate the value of spiral CT examination for diagnosis occult fracture of the patients with negative result of X-ray examination and with high suspicion of fractures,so as to reduce misdiagnosis.

METHODSFrom January 2007 to June 2010, 31 patients with ankle trauma performed spiral CT examination, including 18 males and 13 females, ranging in age from 21 to 67 years, with a mean of 35 years. The main symptoms of the patients included ankle pain, local swelling, obvious tenderness and activity limitation. All the patients had negative results of X-ray examination.

RESULTSThe spiral CT examination revealed 11 patients with fractures, involving a total of 17 points. Single fracture were found in 6 cases,and multiple fractures were found in 5 cases. Among single fractures, the lateral malleolus fracture was found in 1 case, talus fracture was found in 1 case, scaphoid fracture was found in 1 case, the fracture of the base of 5th metatarsal base was found in 1 case and calcaneal fractures were found in 2 cases. Within multiple fractures,internal and lateral malleolus fracture were found in 1 case; medial malleolus, calcaneus and talus fractures were found in 1 case; talus and scaphoid fractures were found in 1 case; the fractures of 1st and 2nd cuneiform bone were found in 1 case; the 2nd and 3rd metatarsal base fracture was found in 1 case.

CONCLUSIONFor the patients with negative X-ray examination and high suspicion of fractures,the spiral CT examination is needed, which could significantly improve the detection rate of occult fractures, and provide imaging basis for clinical treatment and judicial identify.

Adult ; Aged ; Ankle Injuries ; diagnostic imaging ; Female ; Foot Injuries ; diagnostic imaging ; Fractures, Closed ; diagnostic imaging ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Tomography, Spiral Computed ; methods

We cloned the lipoxygenase gene (ana-LOX) from Anabaena sp. PCC 7120 and expressed it in Escherichia coli BL21 (DE3) pLysS. We determined the active site of the recombinant ana-LOX through site-directed gene mutagenesis and obtained the shortest length of the functional gene. Meanwhile, we studied the properties of recombinant ana-LOX after purification. The C-terminal of the Aos (allene oxide synthase)-LOX fusion gene in Anabaena sp. PCC 7120 genome was found belonging to LOXs family by bioinformatics analysis. Further results of site-directed gene mutagenesis confirmed that the active sites of ana-LOX were His197, His202, His369, Asn373and Ile455. The shortest length of functional gene was identified to be 1 254 bp based on the strategy of shortening the gene length gradually. The highest activity of recombinant ana-LOX of 6 750 U/mL could be achieved when constructed to pET-32a vector and expressed at low temperature 16 degrees C. We purified the enzyme by Ni-NTA chelating affinity chromatography, with 60.89% yield and specific activity of 11.4 x 10(4) U/mg. The optimum reaction temperature and pH for ana-LOX were 45 degrees C and 6.0, respectively. Furthermore, the obtained ana-LOX was stable at room temperature. The effect of metal ions on ana-LOX was determined also. Fe2+, Mg2+ Ca2+ could markedly promote the activity of this enzyme whereas Fe3+ and Cu2+ had a strong inhibitory effect on it. Finally, the ana-LOX could improve the microscopical structure of dough. The results of this study will provide a basis for future improvements and food industrial applications of ana-LOX.
Anabaena ; enzymology ; genetics ; Catalytic Domain ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; metabolism ; Lipoxygenase ; chemistry ; genetics ; Metals, Heavy ; chemistry ; Mutagenesis, Site-Directed ; Recombinant Proteins ; chemistry ; genetics

OBJECTIVETo evaluate the performance of computer-assisted imaging system in the detection of cervical squamous intraepithelial lesion and quality-assurance.

METHODSManual PAP screening (n = 140 580) and image-assisted screening (n = 32 885) were compared for the detection rates of squamous cell abnormalities, the atypical squamous cells (ASC) to squamous intraepithelial lesion (SIL) ratio, the positive rates of high risk human papillomavirus (HR-HPV) test in the case of atypical squamous cells of undetermined significance (ASC-US), and the correlation between cytopathology and histopathology.

RESULTSCompared with manual screening, computer-assisted imaging system showed increased overall positive detection by 0.32%, decreased detection of ASC by 0.21%, increased detection of low-grade squamous intraepithelial lesion (LSIL) by 0.22%, increased detection of high-grade squamous intraepithelial lesion or worse (HSIL) by 0.31%, and decreased ASC to SIL ratio from 2.59 to 1.60. Computer-assisted imaging system did not change the HR-HPV positive rate of the patients who were ASC-US, or the coincidence rate between cytopathology and histopathology. Moreover, the productivity of the laboratory operation increased 58.33%.

CONCLUSIONComputer-assisted imaging system significantly increases the overall positive detection rate of cervical SIL, improves accuracy and work efficiency of screening, decreases the ASC/SIL rate, and strengths the quality-assurance of laboratory testing.

Carcinoma, Squamous Cell ; pathology ; Cervical Intraepithelial Neoplasia ; pathology ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; pathology ; Uterine Cervical Dysplasia ; pathology ; Uterine Cervical Neoplasms ; pathology ; Vaginal Smears ; methods

Aptamers are capable of binding a wide range of biomolecular targets with high affinity and specificity. It has been widely developed for diagnostic and therapeutic purposes. Because of unique three dimensional structures and cell-membrane penetration, aptamers inhibit virus infection not only through binding specific target, such as the viral envelope, genomic site, enzyme, or other viral components, but also can be connected to each other or with siRNA jointly achieve antiviral activity. Taking human immunodeficiency virus and hepatitis C virus as examples, this paper reviewed the effects and mechanisms of aptamers on disturbing viral infection and replication steps. It may provide an insight to the development of aptamer-based new antiviral drugs.
Antiviral Agents ; pharmacology ; Aptamers, Nucleotide ; pharmacology ; therapeutic use ; Genome, Viral ; drug effects ; HIV ; drug effects ; HIV Reverse Transcriptase ; metabolism ; Hepacivirus ; drug effects ; genetics ; Humans ; Macular Degeneration ; drug therapy ; Neoplasms ; drug therapy ; Oligodeoxyribonucleotides ; therapeutic use ; RNA, Small Interfering ; pharmacology ; SELEX Aptamer Technique ; Viral Envelope Proteins ; metabolism ; Virus Replication ; drug effects