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1.
Acta Pharmaceutica Sinica ; (12): 2695-2701, 2020.
Article in Chinese | WPRIM | ID: wpr-837522

ABSTRACT

Heavy metals and other harmful elements in traditional Chinese medicines inflict serious damage on public health. Therefore, risk assessment of Chinese raw materials has gained increasing attention. To date, few reports have been published on the health risk assessment of heavy metals and harmful elements in Chinese patent medicines. To gain a comprehensive understanding of heavy metals and other harmful elements in Chinese patent medicines and to establish proper limits, residual Pb, Cd, As, Hg, Cu and Cr in 15 054 samples of 295 drugs was analyzed with regard to distribution and variation between elements and dosage forms. In addition, in accord with procedures including hazard identification, hazard characterization, exposure assessment and risk characterization, basic procedures and specific parameters for risk assessment of heavy metals and harmful elements in Chinese patent medicines were clarified based on the health risk assessment of 14 787 samples and 276 drugs. A method and equation for establishing residual limits is proposed. The results show that content and target hazard quotients (THQs) of the investigated elements in all samples showed a skewed distribution approaching 0. Content of Pb, As, Cu, Hg, Cd or Cr in the samples exceeded 100 mg·kg-1 and the content of Pb, As, or Cu in individual samples exceeded 1 000 mg·kg-1. THQs of 586 samples and four drugs were above 1. We believe that the health risk of Hg, Pb and As in Chinese patent medicines with dosage forms of pill, capsule, tablet and powder, especially those in raw powder preparations, warrant concern.

2.
Herald of Medicine ; (12): 235-237, 2019.
Article in Chinese | WPRIM | ID: wpr-744221

ABSTRACT

Objective To establish limit test method of camphor in Niuhuang qingxin pills (prescription of the bureau) . Methods The separation was performed on an HP-INNOWAX capillary column (0.25 mm×30 m, 0.25 μm) and the column temperature was 110 ℃ . The temperatures of the inlet and the FID detector were 200 and 230 ℃ , respectively. The flow rate of the carrier gas was 1.8 mL·min-1 and the split ratio was 10:1. The injection volume was 1 μL. Results Good linearity was obtained for camphor within the range of 1.65 to 165 μg·mL-1, and the average recovery of low, medium, high concentration were 99.12%, 99.56% and 99.56%, respectively,with RSD were 1.01%,0.69% and 0.38%, respectively. Trace camphor was found in 7 samples out of 38 samples from 9 manufactures. Conclusion The proposed method was accurate, sensitive and simple, and was suitable for quality evaluation and safety control of Niuhuang qingxin pills (prescription of the bureau) .

3.
Acta Pharmaceutica Sinica ; (12): 2074-2082, 2019.
Article in Chinese | WPRIM | ID: wpr-780288

ABSTRACT

We determined a component-target-disease network for Carthamus tinctorius L. and the key compounds, identified by topological analysis, were related to vasculitis, coronary heart and cerebrovascular disease. Based on these compounds, the chromatographic fingerprint of Carthamus tinctorius L. was established. Firstly, 132 compounds were obtained from TCMID and TCMSP databases. Their targets were predicted in the PharmMapp and HemMapper databases. CardioGenBase, Therapeutic Target Database and DisGeNET databases were used to collect targets of vasculitis, coronary heart disease and cerebrovascular disease. The corresponding relationships between component and target protein were established by mapping. Finally, the "component-target-disease" network was built with Cytoscape software. The core network and key nodes were analyzed with the Cytohubba plug-in. The results showed that the 24 key compounds were alpha-tocopherol, adenosine, quinone chalcone pigments such as hydroxysafflor yellow A, safflower yellow, quercetin, kaempferol and other flavonoids, organic acids such as stearic acid, linolenic acid, coumaric acid and cinnamic acid. This resulting chromatographic fingerprint of Carthamus tinctorius L. showed good consistency, and the core chemical compounds obtained by topological analysis of the network of "component-target-disease", could be used as quality control markers. Our research provides a new approach for the identification of quality control indicators in Chinese medicinal materials.

4.
Acta Pharmaceutica Sinica ; (12): 354-359, 2019.
Article in Chinese | WPRIM | ID: wpr-780099

ABSTRACT

Though red yeast rice (RYR) has been used as medicine for centuries, few study has been reported about its biological activities related to traditional medicinal application and marketed RYR showed poor consistency in quality. In this study, with comprehensive investigation of their production processes and field acquisition samples including those from genuine producing area, an ultra performance liquid chromatographic (UPLC) method was firstly established to discriminate RYR for different applications based on their secondary metabolites fingerprint. It was performed on a CAPCELL CORE AQ column (100 mm×4.6 mm, 2.7 μm), with PDA (range: 200-650 nm, extracted: 237 nm) and ELSD detection. The mobile phase used was water (A) and acetonitrile (B) both containing 0.1% formic acid at gradient elution (0-15 min, 50% B→85% B (linear); 15-16 min, 85 % B→50% B (linear) and maintained until 21 min), with a flow rate of 0.5 mL∙min-1. The method established was fully validated in agreement with guidelines of Chinese Pharmacopeia. Common metabolites were found in RYR for same application and the fingerprints of RYR for food coloring or brewing from various manufacturers had similarities above 0.90. Meanwhile, significant differences were observed among the fingerprints for various applications and discrimination could be achieved by principal component analysis (PCA). Lovastatin was absence in RYRs for food coloring or brewing, and the fingerprint of traditional medicinal RYR was similar to that of RYR for brewing. However, standardization was required for RYR containing lovastatin because of their significant differences from various manufacturers in fingerprints and lovastatin content. The results demonstrated the feasibility to discriminate RYR for different applications by the secondary metabolites fingerprint method established in this study, which provides a scientific basis to investigate the relationship between biological activities of medicinal RYR and their corresponding secondary metabolites, and further aid their quality standardization and improvement.

5.
Article in Chinese | WPRIM | ID: wpr-773708

ABSTRACT

Tripterygium wilfordii preparations,with various biological activities such as immunosuppressive,anti-inflammatory and anti-cancer effects,are widely used in the treatment of autoimmune diseases such as rheumatoid arthritis,lupus erythematosus,and nephrotic syndrome. They have definite therapeutic effect,but often cause serious adverse reactions and result in damages to liver,kidney,blood,reproduction,and other systems due to their complex compositions,great toxicity,and narrow margin between the toxic and therapeutic dosages. At present,T. wilfordii preparations produced by different manufacturers exhibit large variations in clinical efficacy and side effects in account of their different chemical compositions and quality fluctuation due to differences in raw materials and production process. However,the existing quality standards are controversial in terms of index components and content limit,which cannot be effectively used for the overall quality control of the preparations. In this paper,the research progress on chemical constituents,quality standard and quality control methods of four T. wilfordii preparations including Tripterygium Tablets,Tripterygium Zongtie Tablets,Tripterygium Shuangceng Tablets and Tripterygium Glycosides Tablets was reviewed,in order to provide ideas and reference for the quality improvement of this type of preparations.


Subject(s)
Drugs, Chinese Herbal , Reference Standards , Quality Control , Tablets , Tripterygium , Chemistry
6.
Article in Chinese | WPRIM | ID: wpr-771514

ABSTRACT

To achieve a comprehensive understanding of heavy metals and harmful elements residues in Niuhuang Qingwei Pills,49 samples from 18 manufactures were collected from 31 provinces in China.Risk assessment and control preparations were applied innovatively in evaluation of exogenous pollution in traditional Chinese Medicine.Determination methods for Pb,Cd,As,Hg and Cu were established by inductively coupled plasma mass spectrometry(ICP-MS).Based on the procedures including hazard identification,hazard characterization,exposure assessment and risk characterization,risk assessment was performed and residual limits for Pb,Cd,As,Hg and Cu in the drug were formulated.The results showed that the hazardous quotients(HQ) of the elements were decreased in the following order:Pb>As>Cu>Hg>Cd,and the total hazardous index(HI) of heavy metals and harmful elements in Niuhuang Qingwei Pills was above 1,implying health risk of the drug.Under the proposed limits,5 elements in the control preparation as well as Cd and Cu in the samples were within the limits range,but the excess rates of Pb,As and Hg in the samples were 12%,12% and 14%,respectively.For the first time,basic steps for risk assessment of Chinese patent medicine were established,which provided model and reference for risk assessment and limit formulation of other drugs.


Subject(s)
China , Drug Contamination , Drugs, Chinese Herbal , Reference Standards , Metals, Heavy , Risk Assessment
7.
China Pharmacy ; (12): 2027-2030, 2019.
Article in Chinese | WPRIM | ID: wpr-817175

ABSTRACT

OBJECTIVE: To provide reference for perfecting the quality standard revision of TCM preparations in medical institutions, and improving the quality standard of TCM preparations in medical institutions. METHODS: By sorting out the quality standards of TCM preparations in Guangdong provincial medical institution preparation specification, this paper summarized the main problems and put forward suggestions for improvement. RESULTS & CONCLUSIONS: There are 897 kinds of TCM preparations included in the Guangdong provincial medical institution preparation specification. The recent quality standards (2017 edition) have been greatly improved compared with those of the first edition (1985 edition); however, there are still some problems in the overall quality control of preparations, quantitative control of indicative components, project specificity, source control of medicinal materials, control of medicinal materials not included in the Chinese Pharmacopoeia and quality standards of the same series of varieties. It is suggested that on the premise of considering both advancement and applicability, the quality control of the whole, local and toxic preparations of TCM hospitals should be strengthened, the detection methods with good specificity and reproducibility should be properly updated, the control of key parameters of production process should be strengthened, the quality standard of TCM preparations in medical institutions should be fully improved, so as to provide the products with safe, effective and controllable in quality products in medical institutions.

8.
Article in Chinese | WPRIM | ID: wpr-801946

ABSTRACT

Objective: To establish an high performance liquid chromatography (HPLC) method for the simultaneous determination of four constituents in Niuhuang Qingwei pill (narirutin,naringin,hesperidin,and neohesperidin), and identify the source of Fructus Aurantii Immaturus. Method: The analysis was performed on a Waters CORTECS C18 column (4.6 mm×50 mm,2.7 μm), with acetonitrile-0.12% formic acid solution as the mobile phase for gradient elution. The flow rate was 0.5 mL·min-1, the detection wavelength was set at 283 nm, and the column temperature was 27℃. Result: 12 batches of Niuhuang Qingwei pills showed the different content of flavonoids as Citrus aurantium and C. sinensis. Narirutin,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 5.47-2 735 ng (r=0.999 6),7.25-3 625 ng (r=0.999 5),8.41-4 205 ng (r=0.999 4) and 8.36-4 180 ng (r=0.999 5),and their average recoveries were 101.3% (n=6,RSD 2.9%),98.0% (n=6,RSD 1.8%),95.9% (n=6,RSD 0.8%) and 96.0% (n=6,RSD 1.1%), respectively. The contents of narirutin,naringin,hesperidin,neohesperidin and total flavonoids were 0.36-1.28,2.66-4.87,1.02-11.07,3.58-6.41,and 7.98-13.34 mg·g-1, respectively. Conclusion: The developed method was simple,accurate and reliable,which can be used to identify the source of Aurantii Immaturus Fructus and simultaneously determine the content of four flavonoids in Niuhuang Qingwei pills. It could provide basic research for quality control and composition comparison of 2 kinds of Niuhuang Qingwei pills, showing more comprehensive indicators and reference value for the quality standard improvement of Niuhuang Qingwei pill.

9.
Acta Pharmaceutica Sinica ; (12): 439-443, 2018.
Article in Chinese | WPRIM | ID: wpr-779894

ABSTRACT

The antioxidant activities of Vleriana jatamansi Jones were investigated and the relationship between the antioxidant effect and the chemical structure was explored. The free radical scavenging test, 2,2-diphenyl-1-picrylhydrazyl (DPPH•), was used to evaluate the antioxidant activities of the extracts of Vleriana jatamansi Jones with 0−100% menthol as extraction solvents. The polar and nonpolar HPLC conditions were conducted to isolate the main chemical compositions. The DPPH• tests were used in analysis of the free radical scavenging activities. Under polar HPLC separation conditions, 5 kinds of compounds were detected:chlorogenic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, hesperidin, and coffeic acid; under nonpolar HPLC separation conditions, acevaltrate, 1β-acevaltrate and valtrate were founded. Chlorogenic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid presented high DPPH• free radical scavenging activities. The results of antioxidant activity suggested that the coffee acyl from chlorogenic acid-like compounds had a high DPPH• free radical scavenging ability. Our investigation indicated that structure of the ortho hydroxyl phenol of chlorogenic acid-like compounds play a significant role in antioxidant activities. In addition, this work can also provide method and theory reference for improving the antioxidant activities of Vleriana jatamansi Jones.

10.
China Pharmacist ; (12): 981-983, 2018.
Article in Chinese | WPRIM | ID: wpr-705643

ABSTRACT

Objective: To establish a GC method for the determination of borneol and isoborneol in Niuhuang Qingxin pills ( pre-scription of the bureau). Methods: A capillary column HP-INNOWAX(30 m×0. 25 mm,0. 25 μm) was used and the column tem-perature was kept at 110 ℃. The temperature of the inlet and the FID was 200 ℃ and 230 ℃, respectively. The carrier gas was N2 and the flow rate of the carrier gas was 1. 8 ml·min-1. The split ratio was 10: 1, and the injection volume was 1 μl. Results: The linear response range of borneol and isoborneol was 0. 01-5. 09 μg·ml-1(r=0. 999 5) and 0. 01-5. 03 μg·ml-1(r=0. 999 1), re-spectively. And the average recovery was 99. 34% and 99. 24% with the RSD of 0. 59% and 0. 62% , respectively (n=6). Conclu-sion: The proposed method is accurate, sensitive and simple, and can be used for the quality control of Niuhuang Qingxin pills (pre-scription of the bureau).

11.
Article in Chinese | WPRIM | ID: wpr-687319

ABSTRACT

Ultra-performance liquid chromatography-evaporative light scattering detection (UPLC-ELSD) fingerprint analysis method was established for quality control of Guci tablets. Chromatographic separation was performed on Waters Acquity UPLC BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm) at 30 °C of column temperature. Acetonitrile-0.1% formic acid solution was adopted as mobile phase for gradient elution. The flow rate was set at 0.3 mL·min⁻¹, and the injection volume was 3 μL. Detection was carried out on an ELSD with a nitrogen pressure of 0.28 MPa, drift tube temperature of 60 °C, and gain of 400. A total of 39 batches of samples produced by six manufacturers were measured by using the above method and the data were analyzed by ChemPattern software. The peak present in more than 75% of the samples was defined as a common peak, and 30 common peaks were determined. Among them, 19 peaks were identified by rapid resolution liquid chromatography/tandem mass spectrometry (RRLC-MS/MS) method, 16 of which were confirmed by reference substances. The similarity of the tested samples was 0.47-0.98, suggesting that the quality of the samples from different manufacturers varied greatly. Furthermore, principal component analysis (PCA) and hierarchical analysis (HCA) were performed to clarify the main different components in samples. The results indicated that there might be some feeding problems about Paeoniae Radix Alba, Notoginseng Radix et Rhizoma, and Clematidis Radix et Rhizoma in a few manufacturers. This study provided some evidences for the overall quality control of Guci tablets, as well as its quality standard improvements.

12.
Article in Chinese | WPRIM | ID: wpr-335801

ABSTRACT

Traditional Chinese medicine (TCM) reference drug is a new form of TCM standard reference substance. The purpose of this guideline is to guide the establishment of the reference drug and standardize its investigation and application in national drug standards. Definition of TCM reference drug was specified and relating guideline and technical requirement were introduced in this paper. Its application in quality control of TCM was analyzed and the developing train was proposed. There is a wide prospect for the application of reference drug in quality control of TCM. Thus it has practical significance to explore and conduct the quality evaluation system by using TCM reference drug as the reference substance.

13.
Drug Evaluation Research ; (6): 1005-1012, 2017.
Article in Chinese | WPRIM | ID: wpr-662852

ABSTRACT

Research progress ofpolysorbate 80 was summarized,including standards,chemical composition and impurity test.Assay methods of polysorbate 80 in preparations were also summarized.In addition,characteristics of the current technologies were discussed.Compositions of polysorbate 80 could be detected directly by spectrophotometry,SEC-ELSD or LC-MS methods.In addition,they could be analyzed indirectly by determing the hydrolysates with HPLC-UV or GC methods.In recent years,remarkable progress has been achieved in chemical composition determination and impurity test ofpolysorbate 80.But assay method of the excipient in related drugs need further study,due to large difference between purities ofpolysorbate 80 raw materials.

14.
Drug Evaluation Research ; (6): 931-934, 2017.
Article in Chinese | WPRIM | ID: wpr-662771

ABSTRACT

Objective To study the toxic effect ofNiuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40,200,400,800,1 200 tg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage,exceeding the normal effective dose).While,there was toxicity to embryo development at the concentration greater than 200 tg/mL.The head and eyes of the embryo were smaller,and the pupils were colorless.Zebrafish embryonic heart malformation was observed,heart rate was weak,blood pool congestion happened and back blood flow was not smooth.The abdominal surface of the embryo was poor in transparency,the tail was twisted,the yolk sac elongation structure became thicker,the notochord became thin,there was no normal embryo movement,and the body length became shorter.After the withdrawal,the abnormal phenotype was not recovered,and severity of the abnormal phenotype was concentration-dependent.It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 tg/mL.Its toxic effect occurs during embryonic organogenesis.And it mainly affects the development of embryonic nervous system,cardiovascular system and bone.

15.
Drug Evaluation Research ; (6): 1005-1012, 2017.
Article in Chinese | WPRIM | ID: wpr-660876

ABSTRACT

Research progress ofpolysorbate 80 was summarized,including standards,chemical composition and impurity test.Assay methods of polysorbate 80 in preparations were also summarized.In addition,characteristics of the current technologies were discussed.Compositions of polysorbate 80 could be detected directly by spectrophotometry,SEC-ELSD or LC-MS methods.In addition,they could be analyzed indirectly by determing the hydrolysates with HPLC-UV or GC methods.In recent years,remarkable progress has been achieved in chemical composition determination and impurity test ofpolysorbate 80.But assay method of the excipient in related drugs need further study,due to large difference between purities ofpolysorbate 80 raw materials.

16.
Drug Evaluation Research ; (6): 931-934, 2017.
Article in Chinese | WPRIM | ID: wpr-660706

ABSTRACT

Objective To study the toxic effect ofNiuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40,200,400,800,1 200 tg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage,exceeding the normal effective dose).While,there was toxicity to embryo development at the concentration greater than 200 tg/mL.The head and eyes of the embryo were smaller,and the pupils were colorless.Zebrafish embryonic heart malformation was observed,heart rate was weak,blood pool congestion happened and back blood flow was not smooth.The abdominal surface of the embryo was poor in transparency,the tail was twisted,the yolk sac elongation structure became thicker,the notochord became thin,there was no normal embryo movement,and the body length became shorter.After the withdrawal,the abnormal phenotype was not recovered,and severity of the abnormal phenotype was concentration-dependent.It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 tg/mL.Its toxic effect occurs during embryonic organogenesis.And it mainly affects the development of embryonic nervous system,cardiovascular system and bone.

17.
Chinese Journal of Pathophysiology ; (12): 1891-1895, 2017.
Article in Chinese | WPRIM | ID: wpr-660153

ABSTRACT

AIM:To investigate the effects of dexmedetomidine (DEX) on acute alcoholic hepatic injury in mice and to explore the possible mechanisms .METHODS:Kunming mice (n=50) were randomly divided into 5 groups (n=10):normal saline control (NS) group, acute alcoholic hepatic injury model (E) group, low-dose (10μg/kg) DEX (E+L) group, medium-dose (50 μg/kg) DEX (E+M) group and high-dose (100 μg/kg) DEX (E+H) group.The animals were sacrificed at 6 h after gavage of alcohol or normal saline .The levels of alanine aminotransferase ( ALT) , as-partate aminotransferase (AST), triglyceride (TG), malondialdehyde (MDA), glutathione (GSH) and superoxide dis-mutase ( SOD) were measured .The livers were removed for evaluation of histological characteristics and determining the content of tumor necrosis factor-α( TNF-α) amd interleukin-1β( IL-1β) in the liver tissues by ELISA .The expression levels of cytochrome P4502E1 (CYP2E1) and nuclear factor -κB (NF-κB) in the liver tissues were evaluated by Western blot.RESULTS:Compared with NS group, the levels of ALT, AST and TG were obviously increased in E group , which were obviously decreased in E +M and E+H groups.Compared with NS group, the levels of TNF-α, IL-1βand MDA were obviously increase in E group , which were obviously decreased in E +M and E+H groups.Compared with NS group, the activity of SOD and the content of GSH were obviously decreased in E group , which were obviously increased in E +M and E+H groups.Compared with NS group, the expression of CYP2E1 and NF-κB was obviously increase in E group , which was obviously decreased in E +M and E+H groups.Compared with NS group , ethanol induced marked liver histo- logical injury, which was less pronounced in E +M and E+H groups.CONCLUSION: DEX has a protective effect on mouse liver with acute alcoholic injury by the involvement in the processes of antioxidation and antiinflammation , and its mechanism may be associated with the inhibition of CYP 2E1 and NF-κB expression.

18.
Chinese Journal of Pathophysiology ; (12): 1891-1895, 2017.
Article in Chinese | WPRIM | ID: wpr-657754

ABSTRACT

AIM:To investigate the effects of dexmedetomidine (DEX) on acute alcoholic hepatic injury in mice and to explore the possible mechanisms .METHODS:Kunming mice (n=50) were randomly divided into 5 groups (n=10):normal saline control (NS) group, acute alcoholic hepatic injury model (E) group, low-dose (10μg/kg) DEX (E+L) group, medium-dose (50 μg/kg) DEX (E+M) group and high-dose (100 μg/kg) DEX (E+H) group.The animals were sacrificed at 6 h after gavage of alcohol or normal saline .The levels of alanine aminotransferase ( ALT) , as-partate aminotransferase (AST), triglyceride (TG), malondialdehyde (MDA), glutathione (GSH) and superoxide dis-mutase ( SOD) were measured .The livers were removed for evaluation of histological characteristics and determining the content of tumor necrosis factor-α( TNF-α) amd interleukin-1β( IL-1β) in the liver tissues by ELISA .The expression levels of cytochrome P4502E1 (CYP2E1) and nuclear factor -κB (NF-κB) in the liver tissues were evaluated by Western blot.RESULTS:Compared with NS group, the levels of ALT, AST and TG were obviously increased in E group , which were obviously decreased in E +M and E+H groups.Compared with NS group, the levels of TNF-α, IL-1βand MDA were obviously increase in E group , which were obviously decreased in E +M and E+H groups.Compared with NS group, the activity of SOD and the content of GSH were obviously decreased in E group , which were obviously increased in E +M and E+H groups.Compared with NS group, the expression of CYP2E1 and NF-κB was obviously increase in E group , which was obviously decreased in E +M and E+H groups.Compared with NS group , ethanol induced marked liver histo- logical injury, which was less pronounced in E +M and E+H groups.CONCLUSION: DEX has a protective effect on mouse liver with acute alcoholic injury by the involvement in the processes of antioxidation and antiinflammation , and its mechanism may be associated with the inhibition of CYP 2E1 and NF-κB expression.

19.
Article in Chinese | WPRIM | ID: wpr-510654

ABSTRACT

BACKGROUND:Recent studies have found that long non-coding RNAs (lncRNAs) can regulate stem cel proliferation and differentiation. But it is unclear that how lncRNA NR_033474 regulate stem cel proliferation and cel cycle. OBJECTIVE:To investigate the effect of lncRNA NR_033474 on the proliferation and cel cycle regulation in C3H10T1/2 mesenchymal stem cel s after the NR_033474 overexpressed by lentivirus, and to study the possible regulation mechanism of NR_033474 on mesenchymal stem cel s. METHODS:LncRNA NR_033474 was cloned into a lentivirus vector. Lentivirus particles were infected into C3H10T1/2 cel s to upregulate the expression of NR_033474. The NR_033474 expression level was detected by real-time PCR. Compared with the empty lentivirus vector, the proliferation of C3H10T1/2 cel s which overexpressed NR_033474 was detected by cel counting assay and cel cycle was detected using flow cytometry. The expression of cel cycle-associated proteins such as CDK1, Cyclin B1, Cyclin D1 and P53 were detected by western blot assay. RESULTS AND CONCLUSION:Compared with the control group, lncRNA NR_033474 in C3H10T1/2 cel s which overexpressed NR_033474 was increased by about 100 times (P<0.01), and the proliferation of C3H10T1/2 cel s was significantly inhibited after NR_033474 overexpression by lentivirus (P<0.05). In addition, flow cytometry showed that C3H10T1/2 cel s overexpressing NR_033474 were arrested in G2/M phase compared to the control group. Western blot showed that the expression levels of CDK1 and Cyclin B1 were downregulated, while there were no changes in Cyclin D1 and P53 expression. To conclude, these findings suggest that the NR_033474 overexpression significantly inhibits the cel growth of C3H10T1/2 cel s, at least in part, through induction of cel cycle arrest at G2/M phase.

20.
Acta Pharmaceutica Sinica ; (12): 439-2016.
Article in Chinese | WPRIM | ID: wpr-779189

ABSTRACT

Free amino acids play a great role in traditional Chinese medicine in injections of animal products, as they may take part in peptide synthesis and exhibit a bioactivity in vivo. However, most of the national standards for drugs and peer-reviewed papers only focus on the total amount of amino acids after peptide hydrolysis. We compare the advantage and disadvantage among high performance thin layer chromatography (HPTLC), pre-column derivatization ultra performance liquid chromatography (UPLC) and ion chromatography. As a result, the HPTLC and pre-column derivatization UPLC are suitable for quality analysis, while there is high matrix influence for ion chromatography analysis. The verified pre-column derivatization UPLC method is utilized in quantitative analysis. 24 kinds of amino acid were detected by this method, and 8 of them were reported for the first time from the injection. The system has high repeatability and accuracy with LOD on the level of pmol·mL-1.

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