To control the quality of Humulus scandens, the quality standard was established in this study. According to the method recorded in the Appendix of Chinese Pharmacopoeia (2010 Edition) , the water and ash inspections were carried out. The component luteoloside and cosmosiin in Humulus scandens were identified and assayed by TLC and HPLC. The results showed a strong characteristics microscopic of Humulus scandens, and trichoromethane-methanol-formic acid (10: 3: 0. 3) as the mobile phase of TLC, the spots at 365 nm with a UV lamp was clear. The 16 batches of samples were analyzed by HPLC with a gradient elution of acetonitrile and phosphate solution (0.2%) at a flow rate of 1.0 mL · min(-1) and detected at 350 nm. The content of luteoloside was 0.015%- 0.651% (average 0.148%); the content of cosmosiin was 0.003%-0.118% (average 0.036%). The linear calibration curve of luteoloside and cosmosiin was acquired in the ranges of 0.011-0.364 g · L(-1) (r = 1.000 0) and 0.003-0.096 g · L(-1) (r = 1.000 0), respectively. The average recovery was 100.5% and 98.5%, respectively. The methods are convenient and reliable, which can be ap- plied for quality assessment of Humulus scandens.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; standards ; Humulus ; anatomy & histology ; chemistry ; Quality Control

Drug Monitoring ; statistics & numerical data ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; Humans ; Research Design ; Serum ; chemistry

It is an important task to lab of clinical microbiology to surveille multi-drag or pan-drug resistant strains,such as penicillin-or macrolide-resistant Streptococcus pneumoniae,methicillin-resistant Staphylococcus aureus(MRSA),3rd generation cephalosporin-or quinolone-resistant Enterobacteriaceae, carbapenem-resistant Pseudomonas aeruginosa or Acinetobacter baumannii,and so on.We must understand characteristics of these resistant strains to guide doctors' empirical therapy of infective diseases.

Objective The aim of our study is to establish and characterize the animal model for au- toimmune myositis.Methods Fifty male SD rats were randomly divided into 2 groups:model group(n=40) and control group(n=10).The model group rats were immunized with muscle homogenate every week for 5 weeks and received an injection of 2?g pertussis toxin at the first and second week.As controls,10 SD rats were injected with an equal volume of normal saline.Tissue specimens from limb skeletal muscles were ob- tained at 1,2,3,4,5 weeks after injection.At the same time,the blood samples were collected,and the level of CPK was measured.Results The model group had significantly elevated serum CPK levels.There were multiple inflammatory lesions in the skeletal muscles.Local degeneration and necrosis of muscle fibers with disappeared transverse striation,mononuelear cell infiltration in the interstitial could be observed.The patho- logic grade was mainly 2a.The infiltrating mononuclear cells were predominantly CD8~+T cells that mainly lo- cated in the endnmysium.MHC classⅠantigen expression on muscle fiber membranes in the model group was upregulated.Conclusion The experimental autoimmune myositis induced by syngeneic skeletal muscle ho- mogenate in SD rat is pathologically similar to human myositis.It can be used as a good model for human myositis and provides the basis for the etiopathology and therapeutical studies.

Objective To determine the expression of HPV16/18,31/33 DNA and p53,p21~(WAF1) and MDM2 proteins in invasive squamous cell carcinoma of cervix (ISCC)and to indicate the significance of them in the occurrence and development of ISCC.Methods Using tissue microarray,in situ hybridization (ISH) and immunohistochemical method,we detected the expression of HPV16/18,31/33 and investigate the expres- sion of p53,p21~(WAF1),MDM2 and proteins in ISCC,CIN and NCE.Results was analysed by SPSS vision 12.5. Results The positive expression rate of HPV16/18,p53,p21~(WAF1),MDM2 in ISCC was markedly higher than in CIN and NCE.We found the difference between HPV31/33 and lymph node transfer.Significant relation- ship was observed between p53 protein expression and histological grade and lymph node metastasis of the cancer.There was positive correlation between the expression of p21~(WAF1) protein and the depth of invasion(P

OBJECTIVETo explore the mechanism of Shenqi compound recipe (SQCR) anti-earlier diabetic artherosclerosis in GK rats.

METHODFour-month specefic pathogen free (SPF) GK rats were divided randomly according to blood glucose level into four groups: model group (5 mL x kg(-1) x d(-1) sterile water), ramipril group (positive control, 1 mg x kg(-1) x d(-1)), SQCR low dosage (0.72 g x kg(-1) x d(-1)) and SQCR high dosage group (2.88 g x kg(-1) x d(-1)) and Wistar rats as normal control group(5 mL x kg(-1) x d(-1) sterile water). GK rats took high-fat diet freely and meanwile were injected N-omega-nitro-L-arginine methyl ester (L-N-AME) intra-peritoneally with the dose of 10 mg x kg(-1) x d(-1) in order to induce earlier diabetic artherosclerosis, while normal control group took regular diet and were injected normal saline intra-peritoneally. In the experiment periods, each group was administrated correspondent substance respectively for 32 d. At the end, sampling blood by abdominal aorta and picking aorta on ice. Determined monocyte chemoattractant protein-1 (MCP-1) concentration by ELISA, messenger ribonucleic acid (mRNA) expression of MCP-1 and peroxisome proliferator-activated receptor gamma (PPARgamma) in aorta by reverse transcriptase PCR (RT-PCR).

RESULTConcentrations of MCP-1 in serum in SQCR low and high dosage groups and the mRNA expression of MCP-1 in SQCR high dosage group were all decreased significantly compared with model group (P < 0.05). The mRNA expression of PPARgamma in SQCR low and high dosage groups all increased compared with model group (P < 0.05 or P < 0.01).

CONCLUSIONInhibiting the mRNA and protein expression of MCP-1 and upregulating the mRNA expression of PPARgamma in aorta might be contribute to SQCR anti-earlier diabetic artherosclerosis in GK rats partly.

Animals ; Aorta ; metabolism ; Astragalus membranaceus ; chemistry ; Atherosclerosis ; etiology ; metabolism ; Chemokine CCL2 ; biosynthesis ; blood ; genetics ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Male ; PPAR gamma ; biosynthesis ; genetics ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo observe the effects of Shenqi Compound (SQC) on the mRNA expression of angiotensin II type 1 receptor (AT1R) in the aorta of Goto-Kakizaki (GK) rats.

METHODSTotally 67 GK rats were randomly divided into 5 groups, i.e., the GK group (n =18), the model group (n =16), the atorvastatin group (n =17), and the SQC group (n =16). Another a normal control group was set up (n =18). The diabetic macrovascular disease model was prepared by adding L-NAME (at the daily dose of 0.10 mg/mL) in drinking water for GK rats. GK rats, except those in the normal control group were fed with high fat diet. Atorvastatin (at the daily dose of 1.60 mg/kg) and SQC (at the daily dose of 1.44 g/kg) were respectively administered by gastrogavage, once daily for 35 successive days. The blood glucose was determined by glucose oxidase method once per week. After 5-week medication, the contents of triglyceride (TG) and total cholesterol (TC) were determined by ELISA. The serum concentrations of angiotensin I (Ang II) were determined by RIA. The mRNA expression of AT1R in the aorta was determined by real-time quantitative reverse transcriptase PCR (RT-PCR).

RESULTSThe blood glucose level was obviously lower in both the atorvastatin group and the SQC group after 4 weeks of medication (P <0.05). Besides, it was significantly lower in the SQC group than in the model group by the end of the 4th week (P <0.05). The concentrations of TG, TC and serum Ang II , and the mRNA expression of AT1R in the aorta were significantly higher in the model group than in the normal control group (P <0.01). After 5-week medication, the concentrations of TG, TC and serum Ang I , and the mRNA expression of AT1 R in the aorta were significantly lower in the atorvastatin group and the SQC group than in the model group (P <0.01, P <0.05). The mRNA expression of AT1R was significantly higher in the SQC group than in the atorvastatin group (P <0.05).

CONCLUSIONSSQC could significantly reduce the levels of blood glucose, TG, TC, down-regulate the mRNA expression of AT1R in the aorta, and decrease the expressions of serum Ang II of GK rats with diabetic macrovascular disease. AT1 R might be one of effective targets of SQC in treating diabetic macrovascular diseases.

Angiotensin II ; blood ; Animals ; Aorta ; drug effects ; metabolism ; Blood Glucose ; analysis ; Cholesterol ; blood ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Male ; RNA, Messenger ; genetics ; Rats ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Triglycerides ; blood

OBJECTIVETo identify mouse acute pneumonia model induced by influenza virus adapted strains (FM1 strain) using RT-PCR, gene clone and sequence analysis and pathological examination of lung tissues.

METHODSAcute pneumonia was induced by intranasal drip of FM1 strain. The lungs were collected after 3, 5 and 7 days. RT-PCR was used to detect the viral load. Amplified PCR products were cloned and sequenced. Pathological and histological changes to the lungs were observed.

RESULTSThere were no abnormalities in the alveoli, alveolar sacs and alveolar septa and no inflammatory cell infiltration was found in normal mice. In the model group, we found disappearance of alveoli, alveolar sacs, alveolar ducts and alveolar septa, thickening of the alveolar septal and bronchiolar walls, and infiltration of inflammatory cells after 3, 5 and 7 days of influenza virus (IV) infection. Compared with the normal group, pathological changes at various time points were significantly increased (P<0.01). Viral nucleic acid can be detected in the lung tissue of the model group at various time points, and the pathological changes of the lung tissue were positively correlated with viral load. Sequence analysis demonstrated that there was 99.1% consistency between RT-PCR products of lung tissues in the model group and the known IV cDNA sequence (P<0.01).

CONCLUSIONSGene clone and sequence analysis may be used to identify acute mouse pneumonia model induced by FM1 strain.

Acute Disease ; Animals ; Base Sequence ; DNA, Complementary ; chemistry ; Disease Models, Animal ; Female ; Influenza A Virus, H1N1 Subtype ; Lung ; pathology ; Male ; Mice ; Molecular Sequence Data ; Pneumonia, Viral ; etiology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

Though all the marketed drugs of dipeptidyl peptidase IV inhibitors are structurally different, their inherent correlation is worthy of further investigation. Herein we rapidly discovered a novel DPP-IV inhibitor 8g (IC50 = 4.9 nmol.L-1) which exhibits as good activity and selectivity as the market drugs through scaffold hopping and drug splicing strategies based on alogliptin and linagliptin. This study demonstrated that the employment of classic medicinal chemistry strategy to the marketed drugs with specific target is an efficient approach to discover novel bioactive molecules.
Dipeptidyl-Peptidase IV Inhibitors ; chemical synthesis ; chemistry ; Drug Design ; Drug Discovery ; methods ; Humans ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; Linagliptin ; chemical synthesis ; chemistry ; Molecular Structure ; Piperidines ; chemical synthesis ; chemistry ; Structure-Activity Relationship ; Uracil ; analogs & derivatives ; chemical synthesis ; chemistry

This study aims to investigate the genetic characteristics of BZLF1 gene and its promoter Zp of the epidemic strains in children with primary Epstein-Barr virus (EBV)-associated diseases. Total DNA was extracted from the peripheral blood of 134 children with EBV-associated infectious mononucleosis (EBV-IM) and 32 children with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) who were admitted to Beijing Children's Hospital from 2006 to 2011. The EBNA3C, BZLF1, and Zp genes were amplified by PCR assay. Typing of EBV was performed according to the size of the amplification product of EBNA3C gene; the amplification products of BZLF1 and Zp genes were subjected to direct sequencing, and sequence analysis was performed using BioEdit 7. 0. 9. The results were as follows: (1) EBV-1 was present in 140 samples (97.2%, 140/144) and EBV-II in 4 samples (2.8%, 4/144). (2) Three BZLF1 genotypes and their 12 subtypes (including 6 newly found subtypes) were detected in this study; there were no significant differences in the frequencies of BZLF1-A and BZLF1-B between the children with EBV-IM and EBV-HLH (P = 0.083); BZLF1-A1 was the dominant genotype in children with EBV-associated diseases; t BZLF1-A mostly had three 29-bp repeats in the first intron of BZLF1 gene, and BZLF1-B mostly had 30-bp repeats (P = 0.000), with the number of repeats varying from 1 to 13. (3) Four Zp genotypes were detected in this study, including Zp-P, Zp-V3, Zp-V4, and Zp-V1; there were no significant differences in the frequencies of these Zp genotypes between children with EBV-IM and EBV-HLH (P = 0.272, 0.252, 1.0, and 1.0, respectively). (4) The linkage analysis of BZLF1 gene and its promoter Zp showed that BZLF1-A1 was highly associated with Zp-V3 (P = 0.000), while BZLF1-B4 with Zp-P (P = 0.000); EBV-I + BZLF1 A1 was highly associated with Zp-V3 (P = 0.000), while EBV-I+BZLF1-B4 with Zp-P (P = 0.000). The conclusions are as follows: (1) BZLF1-A1 is the dominant genotype in children with EBV-associated diseases; there are mostly 29-bp repeats in the first intron of BZLF1 gene for BZLF1-A genotype and 30-bp repeats for BZLF1-B genotype. (2) Zp-P and Zp-V3 are dominant Zp genotypes of EBV in children, which shared similar detection rates. (3) BZLF1-A1 is highly associated with Zp-V3, while BZLF1-B4 with Zp-P; EBV-I+BZLF1-A1 is highly associated with Zp-V3, while EBV-I+BZLF1-B4 with Zp-P.
Child ; Child, Preschool ; China ; epidemiology ; Epstein-Barr Virus Infections ; epidemiology ; virology ; Female ; Genotype ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Infant ; Infant, Newborn ; Introns ; genetics ; Male ; Promoter Regions, Genetic ; genetics ; Repetitive Sequences, Nucleic Acid ; genetics ; Trans-Activators ; genetics