1. Standard update and the quality control system of traditional chinese medicines
Chinese Pharmaceutical Journal 2015;50(20):1752-1753
Unscramble the updated and revised standards of Traditional Chinese Medicines in the Chinese Pharmacopoeia of 2015 edition. Reflect the progress in the quality control of TCMs, and make a kind of help to understand and implement the monographs of the new upcoming Pharmacopoeia.
2.Effects of abnormal nutritional supply during pregnancy on adult hormone resistance in rats
Wei-Wei SONG ; Tao SHANG ; Hong-Bo LIU ; Shi-Mei ZHANG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(01):-
Objective To discuss the effects of abnormal nutritional supply during pregnancy on the adult insulin and leptin resistance in rats.Methods We established the pregnant rat models given either low protein or high nutrition diet,and normal diet group served as control.There were 12 pregnant rats in each group.After vaginal delivery,the pups birth weight were measured.The randomly selected small for gestational age(SGA)from low protein diet group,large for gestational age(LGA)pups from high nutrition diet group and normal birth weight pups from control group were studied at both 4 weeks and 12 weeks after being born.Each group contained 36 rats.The insulin and leptin level were measured by the method of ELISA,and insulin sensitive index were calculated respectively.Results The pups of mothers served with low protein showed obviously lower birth weight than those mothers with normal diet(P0.05),while the weight of fat around kidney,the ratio of fat and body weight(FW/BW)were(0.36?0.14)g,6.5?0.3,which were higher than those of control (0.19?0.13)g,3.4+O.3(P
3.Study on the detecting methods of the imported materia medica--olibanum.
Shang-mei SHI ; Jin-gai TIAN ; Bao-qin WANG
China Journal of Chinese Materia Medica 2002;27(3):170-173
OBJECTIVETo analyse the chemical components of the essential oil of Gum olibanum somalilnds and Gum olibanum Ethiopia, and to set up determination methods of their main components.
METHODTwo kinds of essential oil are identified by GC-MS, and assayed by Gas chromatography, using SE-54 as the packing material (column 2.1 m x 3.2 mm), with column temperature starting from 80 degrees C, holding for 1 min, and then rising at the rate of 15 degrees C per minute to 170 degrees C.
RESULT40 kinds of chemical compounds in the essential oil of Gum olibanum somalilnds and 22 kinds of those of Gum olibanum Ethiopia were identified by GC-MS, the main component in the essential oil of Gum olibanum somalilnds being alpha-pinene, and the main one of Gum olibanum Ethiopia being Octyl acetate 17 batches of samples were determined with the linear range of alpha-pinene being 0-10.80 micrograms, the correlation coefficient being 0.9995, the recovery being 98.16%, RSD being 1.83%; the linear range of Octyl acetate being 0-10.32 micrograms, the correlation coefficient being 0.9996, the recovery being 99.56%, and RSD being 1.36%.
CONCLUSIONThis study can be used for the setting up of the specification of Olibanum.
Acetates ; analysis ; Boswellia ; chemistry ; Materia Medica ; standards ; Monoterpenes ; analysis ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control ; Species Specificity
4.Inflammatory reaction changes with aging in kidneys of human TIMP-1 transgenic mice
Xue-Guang ZHANG ; Xiang-Mei CHEN ; Quan HONG ; Xi-Yao SHANG ; Suo-Zhu SHI ; Zhong YIN ; Guang-Yan CAI
Chinese Journal of Geriatrics 2003;0(12):-
Objective To explore the role of tissue inhibitor of metalloproteinase-1(TIMP-1) during renal senescence by using human TIMP-1 transgenic mice.Methods Renal histological changes of wild type mice and transgenic mice at the age of 3,12,24 months were observed by periodic acid-schiff(PAS)staining of paraffin sections.The numbers of F4/80 positive cells were detected by immunofluoreseence.The protein expressions of TIMP-1,TIMP-2,matrix metalloproteinase(MMP)-9,MMP-2,intercellular adhesion molecule-1(ICAM-1),transforming growth factor?1(TGF-?1),collagenⅢand collagenⅣwere detected by Western blot.The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography respectively.Results Focal renal fibrosis was found in two genotypes with aging.At the age of 24 months,compared with wild type,in kidneys of transgenic type,the expressions and activities of gelatinases were dowregulated (MMP-2:2.08?0.20 vs.3.39?0.43;MMP-9:4.02?0.82 vs.6.72?1.40,all P<0.05);the expressions of collagenⅢ,collagenⅣ,ICAM-1,and TGF-?1 were upragulated(0.72+0.11 vs.0.57?0.09;0.84?0.13 vs.0.6?0.11,0.72?0.12 vs.0.53?0.07; 0.69?0.12 vs.0.45?0.09,all P<0.05),and the numbers of F4/80 positive cells were increased (18.8?4.4 vs.12.7?3.6,P<0.05)with the upregulated expression and activity of TIMP-1(1.10?0.18 vs.0.62?0.09;50.75?7.25 vs.20.64?3.50,P<0.05).Conclusions TIMP-1 could promote age-related renal fibrosis through enhancing inflammation reaction by ICAM-1 upregulation.
5.Development of chromatographic technologies for the quality control of Traditional Chinese Medicine in the Chinese Pharmacopoeia
Ming-Rui SHEN ; Yi HE ; Shang-Mei SHI
Journal of Pharmaceutical Analysis 2021;11(2):155-162
As an important branch of medicine,Traditional Chinese Medicine(TCM)has been applied for the treatment of diseases for thousands of years in China and other countries in East Asia.The Chinese Pharmacopoeia(ChP)is a drug code formulated by the Chinese government,and it includes a special volume for the monographs of TCM,which plays an important role in ensuring the quality of drugs.The use of quality control technology has always been a complex and important factor in TCM.Owing to the chemical diversity of TCM,chromatography technology has been proven to be a comprehensive strategy for the assessment of the overall quality of TCM and has become the main analytical method in the ChP.This article provides an overview of the classical and modern chromatographic technologies applied in the ChP,and summarizes the advantages and disadvantages of each technique in the TCM monographs.In 2020,the new edition of the ChP(the 2020 edition)has been implemented at the end of 2020.This paper also contains a brief introduction about the application of chromatographic technologies in the new edition of the ChP.
6.Inhibitive effects of anti-oxidative vitamins on mannitol-induced apoptosis of vascular endothelial cells.
Kai-yu PAN ; Mei-ping SHEN ; Zhi-hong YE ; Xiao-na DAI ; Shi-qiang SHANG
Journal of Zhejiang University. Science. B 2006;7(10):825-829
OBJECTIVEStudy blood vessel injury and gene expression indicating vascular endothelial cell apoptosis induced by mannitol with and without administration of anti-oxidative vitamins.
METHODSHealthy rabbits were randomly divided into four groups. Mannitol was injected into the vein of the rabbit ear in each animal. Pre-treatment prior to mannitol injection was performed with normal saline (group B), vitamin C (group C) and vitamin E (group D). Blood vessel injury was assessed under electron and light microscopy. In a second experiment, cell culture specimen of human umbilical vein endothelial cells were treated with mannitol. Pre-treatment was done with normal saline (sample B), vitamin C (sample C) and vitamin E (sample D). Total RNA was extracted with the original single step procedure, followed by hybridisation and analysis of gene expression.
RESULTSIn the animal experiment, serious blood vessel injury was seen in group A and group B. Group D showed light injury only, and normal tissue without pathological changes was seen in group C. Of all 330 apoptosis-related genes analysed in human cell culture specimen, no significant difference was seen after pre-treatment with normal saline, compared with the gene chip without pre-treatment. On the gene chip pre-treated with vitamin C, 45 apoptosis genes were down-regulated and 34 anti-apoptosis genes were up-regulated. Pre-treatment with vitamin E resulted in the down-regulation of 3 apoptosis genes.
CONCLUSIONVitamin C can protect vascular endothelial cells from mannitol-induced injury.
Animals ; Antioxidants ; pharmacology ; Apoptosis ; Endothelial Cells ; cytology ; pathology ; Gene Expression Regulation ; Humans ; Mannitol ; chemistry ; pharmacology ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; chemistry ; Oxidation-Reduction ; Rabbits ; Vitamins ; metabolism
8.Analysis of VP7 serotype of rotavirus in infantile diarrhea in Hangzhou area from 2001 to 2003.
Mei-qin TONG ; Li-qin CHEN ; Shi-qiang SHANG ; Zheng-yan ZHAO
Chinese Journal of Pediatrics 2005;43(8):595-598
OBJECTIVETo investigate molecular epidemiologic features of rotavirus (RV) infection in infantile diarrhea in Hangzhou area.
METHODSStool specimens of 683 infants with suspected acute viral enteritis in the autumn and winter of 2001 - 2003 were collected. RV (group A) was detected by using latex agglutination test (LAT). VP7 serotype (G) positive specimens were detected by using enzyme linked immunosorbent assay (ELISA) and then the RNA of the virus was determined with reverse transcription polymerase chain reaction (RT-PCR). cDNA of VP7 gene fragment was sequenced by automatic gene analyzor (ABI3730) and compared with the RV VP7 gene sequences stored in Genebank.
RESULTSRV was detected in 297 of 683 (43.5%) specimens by LAT. The highest frequency of RV (group A) detected was 52.9% (228/431) in patients aged 7 - 18 months. The prevalent serotypes were G1 (36.7%, 109/297) and G3 (30.9%, 92/297), followed by mixed type (11.8%, 35/297), untyped (9.4%, 28/297), G4 (7.1%, 21/297) and G2 (4.0%, 12/297). The prevalent serotypes seen each year were different. G1 (54.9%, 45/82) was the major serotype in 2001 followed by G3 (14.6%, 12/82). In 2003, the major serotype was G3 (43.0%, 63/146) and followed by G1 (29.5%, 43/146). The reliability of ELISA was confirmed by RT-PCR, gene sequencing and homology analysis.
CONCLUSIONThe main prevalent serotypes of VP7 of rotavirus were G1 and G3. The dominant serotypes of rotavirus varied in Hangzhou area from 2001 to 2003.
Antigens, Viral ; classification ; genetics ; metabolism ; Capsid Proteins ; classification ; genetics ; metabolism ; Child, Preschool ; China ; epidemiology ; Diarrhea, Infantile ; epidemiology ; virology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant ; Latex Fixation Tests ; Male ; Molecular Sequence Data ; Prevalence ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; Rotavirus Infections ; complications ; epidemiology ; virology ; Serotyping
9.Rapid diagnosis of neonatal sepsis by 16SrRNA genes PCR amplification and genechip hybridization.
Mei-qin TONG ; Shi-qiang SHANG ; Yi-dong WU ; Zheng-yan ZHAO
Chinese Journal of Pediatrics 2004;42(9):663-667
OBJECTIVETo explore a method for rapid diagnosis of sepsis in newborn infants.
METHODS(1) The primers and oligonucleotide probes were designed and synthesized based on the sequences of bacterial 16SrRNA gene. The gene chip was prepared through the probes printed onto special glass slides. The gene chip included 18 special probes: universal probe 1, universal probe 2, Gram positive bacterial probe, Gram negative bacterial probe 1, Gram negative bacterial probe 2, Staphylococcus aureus, coagulase negative staphylococcus (CoNS) 1, CoNS 2, Escherichia coli, Hemophilus influenzae, Listeria monocytogenes, Streptococcus pneumoniae, Streptococcus agalactiae, Bacteroides fragilis, Bacillus, Meningococcus, Corynebacterium, Propionibacterium; (2) Blood specimens from 285 cases of suspected septicemia were cultured and bacterial 16S rRNA gene was detected separately; DNA isolated from blood specimens and cerebrospinal fluid was amplified by PCR, and PCR products were hybridized with the probes on the gene chips. Hybridization results were scanned and read by laser-scanner.
RESULTS(1) Of the 285 cases, 17 were positive by PCR and the positive rate (5.96%) was significantly higher than that of blood culture (2.81%) (P < 0.01). When blood culture was taken as control, the sensitivity of PCR was 100% and Specificity was 96.75%, the index of accurate diagnosis was 0.968. (2) The 17 specimens which showed positive results by PCR were further hybridized on the gene chip. All were positive by universal probes. Among all of them, 5 were positive by E. coli probe; 4 were positive by Staphylococcus epidermidis; two were positive by Bacillus and Propionibacterium probes, separately; 4 were positive by CoNS. The 8 specimens which showed positive results by both PCR and blood culture, the result of gene chip hybridization coincided with the result of blood culture.
CONCLUSIONDetection of the bacterial 16SrRNA genes in clinical specimens by gene chip hybridization technology can diagnose neonatal septicemia rapidly. This method has higher sensitivity and specificity than blood culture or other methods and can provide a rapid way for the etiological diagnosis of neonatal septicemia. Therefore the genechip method may be valuable and practical in early diagnosis of neonatal septicemia.
Genes, rRNA ; Humans ; Infant, Newborn ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Sepsis ; diagnosis ; Time Factors
10.Establishment and clinical application of a new real time PCR assay for simultaneous detection of human herpesvirus-6A and human herpesvirus-6B.
Mei-Ting CAI ; Yi-Dong WU ; Xiu-Jing WU ; Shi-Qiang SHANG
Chinese Journal of Pediatrics 2009;47(7):527-531
OBJECTIVEHuman herpesvirus 6 (HHV-6) isolates are classified into two variants, HHV-6A and HHV-6B, based on distinct genetic, antigenic and biological characteristics. HHV-6 has been associated with encephalitis in children recently. This study aimed to establish a real time PCR assay for simultaneous detection of the two subtypes of HHV-6, and apply this new assay to children with suspected encephalitis, then analyze the relationship between the infection with HHV-6 and encephalitis in children.
METHODThe universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene (U38) of HHV-6. The 5' end of the probes for HHV-6A and HHV-6B was labeled with the fluorescein reporter tetrachloro-6-carboxyfluorescein and 6-carboxyfluorescein (6-FAM), separately, while the 3' end were quenched with 6-carboxy-tetramethylrhodamine. The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established. Then, the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 10(9) to 10(0) copies/microl, together with controls were used for testing both sensitivity and specificity of the real time PCR assay. The cerebrospinal fluid (CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.
RESULTBoth HHV-6A (strain ZJ-159) and HHV-6B (strain GS) were positive on the real time PCR assay. There were no cross-reaction with herpes simplex virus type 1, type 2 (HSV-1, HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), hepatitis B virus, Staphylococcus aureus, Mycoplasma pneumoniae and human DNA. A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6. The sensitivity threshold was 10 copies/microl for the real time PCR. HHV-6 positive rate by the real time PCR assay was 4.72% (21/445), including 4 cases with HHV-6A infection, 16 cases of HHV-6B infection and 1 case with mixed HHV-6A and HHV-6B infection. The new PCR assay usually took 2 to 3 hours to provide results.
CONCLUSIONThis new real time PCR assay can simultaneously detect both subtypes of HHV-6, and have high specificity and sensitivity. It will provide an early and sensitive diagnosis of HHV-6 encephalitis in children.
Adolescent ; Child ; Child, Preschool ; DNA Fingerprinting ; DNA Primers ; DNA, Viral ; Encephalitis, Viral ; cerebrospinal fluid ; diagnosis ; virology ; Female ; Fluorometry ; Genotype ; Herpesvirus 6, Human ; genetics ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity