Raltitrexed is a specific inhibitor of thymidylate synthase,and has been confirmed that ralti-trexed has no cross tolerance with 5-fluorouracil.Studies have shown that raltitrexed-based combination chemo-therapy has a beneficial effect on the treatment of advanced gastric cancer,and well tolerance,which is worthy of clinical use.

OBJECTIVETo really realize quality control of Chinese herb purifying process, near-infrared spectroscopy (NIRS) was used not only for fast monitoring quality-control index of the process, but also for fast judgment of absorption endpoint.

METHODThe purification process of Carthamus tinctorius extracts with nonionic macroreticular resin was selected as an example. HPLC was used as the reference method to determine the content of HSYA. Quantitative and qualitative detection modes of purification were developed by NIRS combined with partial least squares (PLS) and moving block of standard deviation (MBSD).

RESULTThe correlation coefficient of the calibration model was 0.999, and the RPD for calibration and validation were above 5, of 5.54 and 5.22, respectively. Based on acquisition spectra, absorption endpoint calculated by MBSD was close to that by HPLC, Only 1 min deviation.

CONCLUSIONThe method mentioned above is proved to be convenient, rapid and nondestructive, and is applicable for fast monitoring the content of HSYA and fast judgment of absorption endpoint in purifying process of C. tinctorius extracts.

Objective:To evaluate the status quo and influencing factors of hyperlipidemia management in patients with contracted family doctor service in the community.Method:The baseline data and blood lipid testing results of 752 hyperlipidemia patients (334 males and 418 females) with contracted family doctor service in Yuetan Community Health Service Center from November?2019 to May 2020 were collected. The hyperlipidemic patients were managed by family doctors based on atherosclerotic cardiovascular diseases(ASCVD) riks assessment. The ASCVD risk levels and low-density lipoprotein cholesterol (LDL-C) compliance rate of patients with different general data were compared, and the influencing factors of LDL-C control failure were analyzed by logistic regression.Results:The ASCVD risk assessment showed that among 752 patients there were 172 cases of low risk(22.87%), 167 cases of moderate risk(22.21%),352 cases of high risk(46.81%) and 61 cases of extremely high risk(8.11%). A significant difference was detected in sex,rate of smoking,incidence of overweight or obesity among patients with different ASCVD risk levels ( P<0.05).The overall control rate of LDL-C was 48.8% (367/752), that for low, moderate, high and extremely high risk patients were 83.73% (144/172), 53.89% (90/167), 34.38% (121/352) and 19.67%(12/61), respectively. A significant difference was detected in sex(female: 52.87%, 221/418),age(aged over 80: 58.82%, 110/187), rate of smoking (non-smoking:52.40%, 327/624) and medication compliance (good compliance:52.87%,221/418) between LDL-C control and uncontrol groups (χ2=6.323,11.816,19.022,25.274; P<0.05). Multiple logistic regression analysis revealed that male gender ( OR=1.800,95% CI:1.325-2.419), smoking ( OR=2.630,95% CI:1.726-4.007) and poor medication compliance ( OR= 2.179, 95% CI: 1.581-3.003) were independent risk factors for uncontrolled LDL-C levels. Conclusion:Patients with hyperlipidemia have a relatively high risk of cardiovascular diseases, and their blood lipids are not well controlled. The management of blood lipid should be enhanced in patients with chronic diseases, particularly for male patients with smoking and poor medication compliance.

Japanese encephalitis virus (JEV) is a single-stranded and positive-sense RNA, which has a single ORF (open reading frame), encoding a polyprotein precursor. Non-structural protein 3 (NS3) plays an important role in processing the polyprotein precursor and has become an important drug target of flavivirus. In this study, NS2BH-NS3 gene was amplified by PCR and subcloned to the prokaryotic expression plasmid, resulting pET30a-NS2BH-NS3. The fusion protein was expressed in Escherichia coli BL21 (DE3) in soluble form after induction by Isopropyl beta-D-1-Thiogalactopyranoside (IPTG). The recombinant protein was purified by Ni-NTA affinity column. Then a fluorescence resonance energy transfer (FRET) method was used to determine enzymatic activity and the assay conditions were optimized. After screening 113 compounds, we found two compounds inhibiting the activity of NS2BH-NS3. This study provides a convenient and cost-effective method for screening of JEV NS3 protease inhibitor.
Encephalitis Virus, Japanese ; enzymology ; Escherichia coli ; metabolism ; High-Throughput Screening Assays ; Protease Inhibitors ; chemistry ; RNA Helicases ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Serine Endopeptidases ; metabolism ; Viral Nonstructural Proteins ; metabolism

Objective To investigate the effects of sericin pretreatment on the expression of extracellular matrix(ECM) associated protein in diabetic nephropathy(DN) rats' kidney.Methods Sixty six male SD rats were randomly divided into 3 groups(n=12):normal control group,DN model group and sericine pretreatment group.DN rats model in model group and sericine pretreatment group were established by intraperitoneally injection of streptozotocin(STZ).Blood glucose≥16.7 mmol/L was taken as the standard of successful modelization.The rats in sericine pretreatment group were lavaged with sericine(2.4 g·kg~(-1)·d~(-1)) for 35 days before injecting STZ.The enzymic method was used to measure the blood glucose.Type Ⅳ collagen(cⅣ)and laminin(LN)content in the serum were detected by ELBA.The expression of transforming growth factor-β_1,(TGF-β_1)and tissue inhibitors of maprix metalloproteinase-1(TMP-1) protein in the kidney was observed by immunohistochemical staining.The expression of Smad 3 protein in the kidney was detected by Western blot.Results Compared with normal control rats,the blood glucose,cⅣ and LN content in the serum,TGF-β_1,TIMP-1 and Smad 3 expression in the kidney of the model group rats increased obviously(P<0.01).The blood glucose,cⅣ and LN content in the serum,TGF-β_1,TMP-1 and Smad3 expression in the kidney of rats in sericine pretreatment group were significantly lower than those of the rats in model group(P<0.01).Conclusion Sericin pretreatment can inhibit the activation of TGF-β/Smad 3 signal pathway in the kidney of DN rats,and prevent the decrease of MMPs activity induced by up-regulation of TIMP-1.So sericin can prevent accumulation of ECM and glomerulosclerosis during DN,and has satisfactory apotropaic effects on the development of DN.

The Expert Consensus on the Diagnosis and Therapy of myelodysplastic syndrome (MDS) (2014) will be interpreted in this paper focusing on whether it is scientific and reasonable.Some advises and views will be put forward,hoping that it will be useful to improve the diagnostic and therapeutic ability on MDS for clinicians in our country.

Objective: To establish a high performance liquid chromatography-charged aerosol detector (HPLC-CAD) method in order to obtain fingerprint profile and achieve the amount of isomer, oleanolic acid, and ursolic acid from Verbena officinalis. Methods: The method was developed by a C30 (150 mm × 2.1 mm, 3 μm) column and a linear gradient elution. Acenitrile-0.2% acetic acid was used as mobile phase. The flow rate was 0.3 mL/min; The injection volume was 10 μL and the column temperature was maintained at 20 ℃. Detector was Corona Ultra CAD with 40 oC of nebulization temperature. Results: Seventeen common peaks were observed from the fingerprint profiles of five different area samples. The relative standard divisions (RSDs) of retention time were less than 0.35%. The relative peak areas of the common peaks were in the range of 3.3%-44.0%. The contents of oleanolic acid were ranging from 0.097% to 0.136%, the contents of ursolic acid were ranging from 0.257% to 0.478%, the total contents were ranging from 0.354% to 0.478%. Conclusion: The fingerprint and simultaneous determination of oleanolic acid and ursolic acid in V. officinalis is firstly studied by HPLC-CAD. This method is simple and with good repeatability, which can be used for the quality control of V. officinalis.

Objective: To establish an accelerated solvent extraction (ASE)-charged aerosol detector (CAD) method for simultaneous detection of jujuboside A, jujuboside B, spinosin, and betulinic acid in Zizyphi Spinosae Semen. Methods: The orthogonal design was applied to optimize the extraction parameters of the ASE system. The target compounds were detected by HPLC-CAD with the parameters as follow: Thermo Syncronis C18 (100 mm × 3 mm, 3 μm) column, a gradient elution program with acetonitrile-water as mobile phase at a flow rate of 0.5 mL/min, the column temperature was kept at 40℃. Detector was Corona Ultra CAD with 35℃ of nebulization temperature. Results: Optimization of the ASE parameters with orthogonal design greatly improved the extraction efficiency; All the target compounds could be simultaneously determined in a single run. Good linear relationships (0.998 3-0.999 6) and high relative recoveries were 98.46%-102.02%. Conclusion: The method is rapid, simple, accurate, and thus could be used for the quality control of Zizyphi Spinosae Semen.

Objective To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific for mouse Bcl-2-assoeiated athanogene 1 (BAG-1) and to observe their inhibitory effects on the expression of BAG-1 gene in mouse melanoma B16FI0 ceils. Methods Plasmids named pRNAT-U6.1/Neo-BAG-1, were designed and constructed to target the mouse BAG-1 mRNA coding region. LipofectaminTM 2000 was used to transfect plasmids into BI6F10 cells. Negative plasmid-transfected and tmtransfected B16F10 cells served as negative and blank controls respectively. Forty-eight hours following transfection, G418 was used to select the resistant cells. The mRNA and protein expression of BAG-1 gene was measured by reverse transcription-PCR and Western blot respectively about 1 month after the transfection. Results The eukaryotic expression plasmids, pRNAT-U6.1/Neo-BAG-1, were constructed, and verified by restriction enzyme digestion and DNA sequencing. The transfection rate in B16F10 cells was 20% -30%. Compared with the blank control, the mRNA and protein expression of BAG-1 in BI6FI0 cells was significantly inhibited by BAG-1 shRNA (both P<0.05), and the inhibition rates were (77±4)% and (62 ±2)%, respectively. Conclusions These results indicate that the eukaryotic expression vectors containing shRNA against BAG-1 gene, pRNAT-U6.1/Neo-BAG-1, are successfully constructed, and can significantly inhibit the expression of BAG-1 gene in mouse melanoma B16F10 cells.

To ensure the quality and safety of Panax notoginseng, a method for the simultaneous determination of 10 mycotoxins in Panax notoginseng was developed using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with acetonitrile and purified by HLB multifunction cleanup column. The separation was performed on a Phenomenex Kinetex XB-C18 column by gradient elution using methanol and 5 mmol·L(-1) ammonium acetate as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The linear relationships of the 10 mycotoxins were good in their respective linear ranges. The correlation coefficients (r) ranged from 0.9981 to 1.0000. The LOQs of the 10 mycotoxins were between 0.15 and 8.6 μg·kg(-1). The average recoveries ranged from 73.8% to 107.0% with relative standard deviations (RSDs) of 0.10%-10.9%. The results demonstrated that the proposed method was sensitive and accurate, and suitable for the mycotoxins quantification in Panax notoginseng.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drug Contamination ; Mycotoxins ; analysis ; Panax notoginseng ; chemistry ; Tandem Mass Spectrometry