Objective: To develop an assay for the determination of cinnamaldehyde in Cortex Cinnamomi and Shentai Plaster. Methods: Hypersil C 18 column (200?4.6mm, 5?m) was used. The mobile phase consisted of a mixture of methanol acetonitrile water tetrahydrofuran (25∶15∶55∶5) with a flow rate of 0.8mL?min -1 . The detection wavelength was at 285 nm. Results: The calibration curve was linear in the range of 32～320ng for cinnamaldehyde ( r =0.9995). The average recoveries of cinnamaldehyde in Cortex Cinnamomi and Shentai Plaster were 97.8% (RSD=3.0%) and 92.2% (RSD=2.9%), respectively. Conclusion: This HPLC method is simple, rapid and accurate, and it can be used in the control of product quality and preparation process.

Objective:To explore the value of clinical pharmacist in clinical treatment through the pharmaceutical care on a patient with Japanese B encephalitis complicated with pulmonary infection. Methods:Clinical pharmacist participated in the whole treatment process and gave suggestions on the selection,dosage and course of the drugs prescribed for anti-virus, anti-epilepsy and anti-infection by focusing on the drug interactions and adverse reactions. Results:The treatment course of the patient was smooth, and the pathoge-netic condition was brought under control gradually while no obvious adverse drug reactions occurred. Conclusion:The work of clinical pharmacist can help to optimize treatment programs to ensure the safety and effectiveness of medication.

Objective:To explore the value of clinical pharmacists in the clinical treatment of a patient with Guillain Barré syn-drome through pharmaceutical care. Methods:Clinical pharmacists participated in the entire treatment process and gave rational sug-gestions about the selection,dosage and course of the drugs,such as glucocorticoid,immunoglobulin and antibiotics. Meanwhile,clini-cal pharmacists focused on the adverse drug reactions. Results:The patient′s condition was controlled and improved gradually. During the hospitalization,the patient developed skin rash and abnormal liver function,while the symptom was improved after the anti allergy and liver protection treatment,and no obvious effect on the primary disease was shown. Conclusion:Clinical pharmacists can help to optimize treatment regimen in order to ensure the safety and effectiveness of patients’medication.

With anti-myc and ras oncogene product monoclonal antibodies p62 and p21, 27 cases of squamous epithelial tumors of the skin (SETS) specimens were examined. The results showed that 62.9%(17/27) and 66.7%(18/27) of the specimens expressed p62 and p21 proteins respectively. Thirteen specimens expressed p62 and p21 proteins simultaneously. The results also showed that p62 was mainly seen in the poorly differentiated squamous cell carcinoma specimens, while p21 was mainly in the relatively well-differentiated specimens. The authors consider that myc and ras oncogenes might have different effects on the develpment of the SETS, and their synergy might be associated with the persistant growth of SETS.

Objective To study the effect of batroxobin combined with Yinxingdamo injection on serum endothelin(ET),nitric oxide(NO),superoxide dismutase(SOD),vascular cell adhesion molecule-1(VEGF-1)in patients with sudden deafness(SVCAM-1),to explore the best treatment for patients with sudden deafness.Methods Ninety patients with sudden deafness from June 2014 to June 2015 were recruited as the subjects of this study.The patients in control group were treated with batroxobin and batroxobin and ginkgo dipyridol.The levels of serum ET,NO,SOD,sVCAM-1,time of symptom recovery and the curative effect were observed.Results After treatment,the levels of ET,NO and sVCAM-1 in the observation group were significantly lower than those in the control group [(64.28±5.72)pg/mL vs(67.36±6.31)pg/mL,(43.08±9.53)μmol/(93.24±11.25)NU/mL](P<0.05).The results showed that there was no significant difference between the two groups.The recovery time of tinnitus,auria and vertigo in the observation group were less than those in the control group [(3.86±1.02)d vs(5.97±1.34),(5.03±1.24)d vs(7.37±2.01)d,(8.09±2.10)d vs 9.07±2.37)d](P<0.05).The total effective rate in the observation group was better than that in the control group(93.33％vs 75.55％,P<0.05).Conclusion Batroxobin combined with ginkgo dipyridolum injection can decrease the level of ET,NO and sVCAM-1,improve the level of SOD,and improve the microcirculation of the inner ear.Compared with the single effect of Batroxobin More desirable,worthy of promotion.

Objective To assess the effect of nicotinic acid on hyperphosphatemia in maintenance hemodialysis (MHD) patients.Methods Twenty-six MHD patients with stable disease condition and hyperphosphatemia that could not be successfully controlled by conventional methods were selected.All patients were treated with oral nicotinic acid for 20 weeks.The changes of serum calcium,high sensitivity Creactive protein (hs-CRP),albumin (ALB),aspartate aminotransferase (AST),alanine aminotransferase (ALT),gamma-glutmyltransferase (GGT),total bilirubin (TBIL),derect bilirubin (DBIL),high density lipoprotein cholesferol (HDL-C),low density lipoprotein cholesferol (LDL-C),triacylglycerol (TG),total cholesterol (TC),intact parathyroid hormone,hemoglobin,platelet count,etc were observed before and after treatment.Results Two patients withdrew from this study because.of pruritus and diarrhea after two weeks of oral nicotinic acid.In 24 cases,compared with that before treatment,the serum phosphate,calcium-phosphorus product was decreased [(1.49 ± 0.19) mmol/L vs.(2.13 ± 0.16) mmol/L,(4.40 ± 0.49)mmol2/L2 vs.(5.45 ± 0.36) mmol2/L2] and HDL-C was increased [(1.45 ± 0.14) mmol/L vs.(1.18 ± 0.15)mmol/L] after treatment,and there was significant difference (P ＜ 0.05).There was no significant difference in the serum calcium,iPTH,LDL-C,TG,TC,ALB,AST,ALT,GGT,TBIL,DBIL,hs-CRP,platelet count,hemoglobin between before and after treatment (P ＞ 0.05).Conclusions Nicotinic acid is effective as all adjuvant measure in control of hyperphosphatemia in patients on MHD and can increase HDL-C.It provides a new approach for the treatment of hyperphosphatemia.

AIM: To isolate three kinds of bufadienolides composition from Venenum bufonis in order to investigate their antitumor activity in vitro.METHODS: Alcohol extraction and isolation by Silica gel colume were employed in this study.Their antitumor activities in vitro were evaluated by MTT colorimetric method.RESULTS: With such methods,three kinds of bufadienolides,including Bufalin,cinobufagin,and resibufogenin with purity above 90% were prepared.The in-vitro results showed that three kinds of bufadienolides had prominent inhibition effect on human lung cancer A549 cells,human breast cancer MDA-MB-435 cells in the range of 0.001 ?g/mL ～ 100 ?g/mL.CONCLUSION: Isolation by Silica gel colume can be used to separate three kinds of bufadienolides from Venenum bufonis and its constituents has prominent anti-cancer effects.

BACKGROUND:Preliminary experiments have demonstrated that rat liver homogenate supernatants can induce the morphological changes of human umbilical cord mesenchymal stem cells. However, little is known about whether the induced cells have some phenotypic and functional features of hepatocytes.
OBJECTIVE:To investigate whether human umbilical cord mesenchymal stem cells have some phenotypic and functional characteristic of hepatocytes after being induced by liver homogenate supernatants.
METHODS:Passage 3 human umbilical cord mesenchymal stem cells were used and divided into control group (cells were cultured in basic culture medium) and liver homogenate supernatant group (cells were cultured in liver homogenate supernatants for 3, 5, 7 days). Meanwhile, positive control group (QSG-7701 human liver celllines) and negative control group (simple liver homogenate supernatants) were set up. The protein and mRNA level of hepatocyte markers, alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme, were detected at different time points.
RESULTS AND CONCLUSION:After inducement, the stem cells of fusiform shape began to lose their sharp edges and progressively shrunk, and then they changed into hepatocyte-like cells with the morphous of triangle, polygon and anomalism shape. Compared with the control group, the protein and mRNA level of alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme significantly increased time dependently after inducement with liver homogenate supernatants (P<0.01). This study demonstrated that human umbilical cord mesenchymal stem cells are able to differentiate into hepatocyte-like cells in vitro that possess some functions of liver cells.

BACKGROUND:Previous studies have demonstrated that normal rat liver homogenate supernatant can induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels with partial hepatocyte functions. However, whether fibrotic liver homogenate supernatant can work or how the inducing effect is remains unclear. OBJECTIVE:To investigate the differentiation potential of human umbilical cord mesenchymal stem cels into hepatocytes under the normal liver and fibrotic liver microenvironment in vitro. METHODS:Liver fibrosis was induced in the SD rats by repeated intraperitoneal injections of 3% thioacetamide at a dose of 200 mg/kg body mass, twice a week for 4 weeks, and then fibrotic liver tissues and normal liver tissues were used to prepare liver homogenate supernatants. Passage 3 human umbilical cord mesenchymal stem cels were used and divided into standard control group (cels were cultured in DMEM/F12 with 10% fetal bovine serum), fibrotic liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 50 g/L fibrotic liver homogenate supernatants), normal liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 100 g/L normal liver homogenate supernatants). The morphological changes of the cels in each group were recorded under inverted microscope; the protein levels of CK18, AFP, CYP3A4, CYP2E1, CYP2D6 and TPH2 were evaluated using western blot assay. Furthermore, the concentration of albumin in the cels was measured. RESULTS AND CONCLUSION:After a 7-day inducement, the stem cels in liver homogenate supernatants groups lost their fusiform shape and changed into hepatocyte-like cels with the morphous of round shape. Compared with the standard control group, the hepatocyte-like cels in the two liver homogenate supernatants groups exhibited human hepatocyte biomarkers, CK18 and AFP. The standard control group cels could express a little amount of CYP2E1, while cels in the two liver homogenate supernatants groups could express CYP3A4, CYP2E1, CYP2D6, TPH2. Compared with the standard control group, the expression level of CYP2E1 in the two liver homogenate supernatants groups increased significantly (P < 0.01), and however, the relative levels of CYP3A4, CYP2E1, CYP2D6, TPH2 in the two liver homogenate supernatants groups showed no statistical significance (P > 0.05). At the same time, compared with the standard control group, the concentration of albumin in the two liver homogenate supernatants groups markedly increased (P < 0.01), but there was no difference between the two liver homogenate supernatants groups (P > 0.05). Experimental findings demonstrated that both of normal liver tissue and fibrotic liver tissue microenvironments could induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels. To achieve the same effect, compared with normal liver tissue, fibrotic liver tissue required lower concentrations, suggesting that fibrotic liver tissue microenvironment may be more conducive to differentiation of umbilical cord mesenchymal stem cels into hepatocytes.

Objective To investigate the effects of NB-UVB on the expression of Gadd45α as well as cell apoptosis and cycle of human HaCaT keratinocytes.Methods Cultured HaCaT cells were exposed to various doses (100,200,400 mJ/cm2)of NB-UVB followed by an additional culture of 6,12 and 24 hours,respectively.Reverse transcription PCR and Western blot were performed to detect the mRNA and protein expression of Gadd45α respectively in HaCaT cells,cell counting kit 8 (CCK8)to measure the proliferation of cells,and flow cytometry to determine the cell cycle distribution of HaCaT cells before and after the exposure to NB-UVB.Results Gadd45α was expressed in HaCaT cells.After exposure to NB-UVB of the three doses,the mRNA and protein levels of Gadd45α increased at 6 hours and 12 hours,but declined at 24 hours,and significant changes were observed in HaCaT cells at the three time points after exposure to NB-UVB of the three doses (all P<0.05).The Gadd45α/β-actin mRNA ratio was 1.4360±0.6551.1.8633±0.0979,1.9266±0.1724 in HaCaT cells 12 hours after irradiation to NB-UVB of 100,200 and 400 mJ/cm2,respectively,significantly higher than that in unirradiated cells(0.6000±0.1276,all P<0.05).Also,increased Gadd45α/β-actin protein ratio was noted in HaCaT cells 12 hours after irradiation to NB-UVB of 100,200 and 400 mJ/cm2 compared with unirradiated cells (0.0773±0.0005,0.1936±0.0015,0.2373±0.0015 vs.0.0290±0.0010,all P<0.05).NB-UVB inhibited the proliferation of HaCaT cells in a time-and dose-dependent manner.Flow cytometry showed that irradiated HaCaT cells were blocked in G2 phase of the cell cycle.and the percentage of HaCaT cells in G2 phase was 13.53%±1.03%,17.77%±2.25%,30.03%±4.29%afler exposure to NB-UVB of 100,200 and 400 mJ/cm2,respectively,compared to 9.24%±0.97%in unirradiated cells (all P<0.05).Conclusions The expression of Gadd45α is increased in HaCaT cells after exposure to NB-UVB,and Gadd45α may be involved in the NB-UVB-induced suprression of cell proliferation of and cell cycle arrest in HaCaT cells.