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OBJECTIVE： To provide reference for the establishment of quality standard of Shenhuang liniment. METHODS： Qualitative identification of Sophora flavescens， Phellodendron chinense， Rhei Radix Et Rhizoma and Scutellaria baicalensis in Shenhuang liniment were carried out by TLC according to the method of 2015 edition of Chinese Pharmacopeia （part Ⅳ）. HPLC method was used to determine the contents of matrine and oxymatrine in Shenhuang liniment [Phenomenex Luna NH2 column， mobile phase consisting of acetonitrile-absolute ethanol-3％ phosphoric acid aqueous solution （80 ∶ 10 ∶ 10，V/V/V）， column temperature of 45 ℃， flow rate of 1.0 mL/min， detection wavelength of 220 nm， sample size of 5 μL]. RESULTS： Results of TLC showed that the corresponding spots of the same color were found in the corresponding positions of the chromatograms for test samples and reference substance/substance control； and the spots were clear， retardation factor was moderate， the separation degree was high， and the negative control had no interference. Results of HPLC showed that the linear range of matrine and oxymatrine were 203.60-1 221.60 ng（r＝0.999 4）and 210.08-840.30 ng（r＝0.999 7）， respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.0％ （n＝6）. Average recovery rates were 96.03％ and 100.93％， and RSDs were 2.55％ and 2.69％（n＝9）. CONCLUSIONS： Established method is specific， accurate and reproducible， and can be used for quality control of Shenhuang liniment.
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Cervical cancer is one of the most common gynecological malignancies which seriously threatens the health of women. How to further expand the scope of screening and improve the sensitivity and specificity of screening and optimize screening programs are the key points of cancer prevention and treatment. At present,the screening methods that are commonly used at home and abroad include cervical smear cytology, human papillomavirus ( HPV) detection,molecular biology detection and genetic genetic testing. Cervical smear cytology has high specificity and low sensitivity,the sensitivity of HPV detection is high,but the specificity is low,and the molecular biological detection and genetic testing has high specificity and high sensitivity,but due to the lack of large quantities of clinical samples,and the higher detection cost,they are still in the test study stage.
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AIM To establish the HPLC fingerprints of Cajanus cajan (L.) Millsp.leaves and to determine the contents of orientoside and luteolin.METHODS The analysis of 65％ methanol extract from C.cajan leaves was performed on a 25 ℃ thermostatic Agilent Zorbax SB-C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-1％ acetic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS There were twenty-one common peaks in ten batches of samples (S1-S10),whose similarities were more than 0.950,except for that of S3 (0.516).Orientoside and luteolin showed good linear relationships within the ranges of 0.089 5-3.960 μg and 0.015 5-0.408 μg,whose average recoveries were 99.43％ (RSD =1.32％) and 98.50％ (RSD =0.82％),respectively.The contents of two constituents in the samples from three growing areas (Guangdong,Yunnan and Hainan) showed obvious differences.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of C.cajan leaves.
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AIM:To investigate the role of immunoproteasome subunit β2i in deoxycorticosterone acetate (DOCA)/salt-induced vascular inflammation in mice.METHODS:Wild-type and β2i knockout male mice were used.The right kidney was removed and DOCA pellet was subcutaneously implanted in the mice.The mice were then received 1% NaCl as drinking water for 3 weeks.The total RNA and protein were isolated from thoracic aorta 3 weeks later.The aortic tissues were fixed in formalin, embedded in paraffin and sectioned.Western blot, real-time PCR and immunohistochemistry were performed to detect the expression of β2i, macrophage marker Mac-2, NF-κB, and proinflammatory cytokines IL-1β, IL-6 and TNF-α in thoracic aorta.RESULTS:Compared with sham group, DOCA/salt treatment significantly increased the expression of β2i at mRNA and protein levels, increased the infiltration of macrophages and expression of Mac-2, and upregulated the expression of NF-κB and proinflammatory cytokines including IL-1β, IL-6 and TNF-α in wild-type group, whereas theses effects were markedly attenuated in β2i knockout mice.CONCLUSION:Immuneproteasome subunit β2i is involved in DOCA/salt-induced vascular inflammation through activation of NF-κB signaling in the mice.
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Objective:To establish a method for the separation and determination of ketoprofen enantiomer .Methods:A pre-col-umn derivation RP-HPLC method was used with L-alanine-β-naphthylamine ( L-Ala-β-NA) as the derivation reagent .The RP-HPLC conditions were as follows: a Hypersil ODS-2 column (250 mm ×4.6 mm,5 μm) was applied, the mobile phase was acetonitrile-0.025 mol· L-1 phosphate buffer solution (40∶60, v/v) and the flow rate was 1.0 ml· min-1 , the detection wavelength was set at 245 nm and the column temperature was 25℃.The injection volume was 10μl.Results:Base line separation was achieved for the sep-aration of enantiomer from ketoprofen , and the retention time for S-(+) -ketoprofen and the R-(-) -ketoprofen was 24.2 min and 26.0 min, respectively.Dexketoprofen within the range of 0.025-0.125 mg had a good linear relationship (r=0.998 1) and the aver-age recovery was 90.93%(RSD =4.10%, n=9 ).Conclusion:The method is simple, accurate and reliable, which can be applied in the separation and determination of ketoprofen .
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Objective:To develop a UPLC method for determining four glycosides in microctis folium.Methods:The UPLC deter-mination was performed on an Agilent SB -C18 (3.0 mm ×50 mm, 1.8 μm) colume with 0.05% acetonitrile -phosphate solution as the mobile phase.The detection wavelength was set at 320 nm.The flow rate was 0.3 ml· min-1.The column temperature was 25℃.Results:Narcissoside, isovitexin, kaempferol-3-rutinoside and isorhamnetin-3-O-glucoside all showed good linearity with the re-covery of 98.89%,99.03%, 97.00%and 97.41%, and RSD of 0.41%, 0.84%,0.44%and 0.80%, respectively (n=9).Con-clusion:The method is convenient with good stability , repeatability and sample recovery rate , which provides basis for the quality eval-uation and control of microctis folium.
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Objective To explore the expression of glucose-regulated protein (Grp) 78 in ovarian cancer tissues and its clinical significance.Methods The expression of Grp78 in 60 cases of ovarian cancer tissue,15 cases of ovarian borderline tumor tissue,10 cases of normal ovarian tissue,10 cases of ovarian cyst tissue,was detected by immunohistochemistry,and analyzed the relationship between its expression and clinicopathological features of ovarian cancer.Results The expression of Grp78 in ovarian borderline tumor and ovarian cancer tissue was significantly higher than that in normal ovarian and ovarian cyst tissue[7/15 and 68.3％ (41/60) vs.1/10 and 1/10] (P ＜0.05).The expression of Grp78 was positively correlated with clinicopathological staging and lymphatic metastasis of ovarian cancer (P ＜ 0.05),negatively correlated with histological differentiation (P ＜ 0.05).No correlation with age and ascites (P ＞ 0.05).Conclusions The levels of Grp78 are elevated in ovarian cancer specimens; high expression of Grp78 maybe participate in the occurrence,development,and prognosis of ovarian cancer.Increased expression of Grp78 might be a useful marker for predicting the carcinogenesis and progression of ovarian cancer.
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BACKGROUND:After glucose regulated protein 94 (GRP94) was knockout in model mice of transplanted tumor, cel ular adhesion is terminated, thus stimulating the proliferation of liver-derived cel s and promoting the development of liver cancer. We speculate that GRP94 plays a protective role against liver cancer. OBJECTIVE:To investigate the expression of endoplasmic reticulum molecular chaperone GRP94 mRNA and protein with smal interfering RNA technique in nude mice model of transplantable human ovarian carcinoma, and to explore the effect of GRP94 mRNA and protein expression on the growth of transplanted tumor. METHODS:The gene sequences of human GRP94 were obtained from Gene Bank. psiSTRIKETM/GRP94 was constructed, which is eukaryotic expression vector control ed by the U6 promoter of human RNA polymerase Ⅲ. The transplantable model of human ovarian carcinoma in nude mice was established using human ovarian cancer HO-8910 cel line. The eukaryotic expression vector was transfected into the transplanted tumors in nude mice, and the growth of the tumor was observed. The nude mice models were divided into three groups, specific smal interfering RNA group, non-specific smal interfering RNA group and saline control group. The volumes of the subcutaneous tumor were determined. RT-PCR and immunohistochemistry were used to detect the mRNA and protein expression of GRP94 respectively. RESULTS AND CONCLUSION:The recombinant plasmid of RNA interference specific for GRP94 was successful y constructed. The subcutaneous tumors appeared in al the nude mice 5 days after transplantation. The diameter of subcutaneous tumors was 7-10 mm 14 days after transplantation. The growth of subcutaneous tumors in nude mice with interference specific for GRP94 treatment was significantly decreased as compared with non-specific smal interfering RNA group and control group (P<0.05). The proliferation activity was inhibited by 65.1%. The expression of GRP94 mRNA and protein was significantly down-regulated after treatment of psiSTRIKETM/GRP94 (P<0.01). The transfection of psiSTRIKETM/GRP94 could significantly induce inhibitory effects on the growth of ovarian carcinoma in nude mice, and the underlying mechanism is associated with the down-regulated expression of GRP94 mRNA and protein.
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AIM: To investigate the changes of alkaloid in Rhizorrm Coptidis and Fructus Evodiae before and after combination. METHODS: The content of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride in Rhizoma Coptidis and Fructus Evodiae before and after combination were determined by TLCS.RESULTS : After combination of Rhizoma Coptidis and Fructus Evodiae,contents of berberine hydrochioride,palmatine hydrochloride,jatrorrhizine hydrochloride significantly reduced,among which the reduction of berberine hydrochloride was more obvious.CONCLUSION: In decoction,Fructus Evodiae appears to restrict the alkaloid from Rhizoma Coptidis in order to lower the side effect of Rhizoma Coptidis.
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AIM:To investigate the changes of alkaloid in Rhizoma Coptidis and Fructus Evodiae before and after combination.METHODS:The content of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride in Rhizoma Coptidis and Fructus Evodiae before and after combination were determined by TLCS.RESULTS:After combination of Rhizoma Coptidis and Fructus Evodiae,contents of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride significantly reduced,among which the reduction of berberine hydrochloride was more obvious.CONCLUSION:In decoction,Fructus Evodiae appears to restrict the alkaloid from Rhizoma Coptidis in order to lower the side effect of Rhizoma Coptidis.