Objective To observe the effect and safety of the compound glycyrrhizin combined recombinant human epidermal growth factor in treatment of eczema. Methods 120 eczema patients were selected randomly divided into the treatment group (n=60) and the control group (n=60) ,the treatment group intravenously given compound glycyrrhizin once a day, 60ml combined recombinant human epidermal growth factor analgesic spray, three times a day. The control group were given intravenously compound glycyrrhizin,once a day ,60ml. Two groups of patients were continuous treatment after a month observation curative effect and adverse reaction. Results The effective rate in the treatment group(91. 67%) was higher than the control group(71.67%) ,there was significant difference(x2=6.43,P＜0.05). Compared with control group, VAS of the treatment group was significantly reduced, there is significant difference (P＜0.05). In the same group,within one month after treatment,before treatment and after treatment compared also has difference, there are significant difference (P＜0.05). The two groups had no serious adverse reactions. Conclusion Effect of compound glycyrrhizin combined recombinant human epidermal growth factor in treatment of eczema curative was distinct,which was worth extending.

Aim To observe the effect of ramipril on ischemia-reperfusion induced apoptosis of cardiomyocytes and expression of apoptosis-related genes.Method Male Wistar rats were subjected to 30 min of myocardial ischemia and 120 min of reperfusion.Rats were randomized to receive vehicle or ramipril(1 mg?kg~(-1)) orally 24 h before surgery.Myocardial infarct size was measured using the staining agent 2,3,5-triphenyl tetrazolium chloride(TTC).Cardiomyocyte apoptosis was assessed using terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) assay and confirmed with detection of DNA ladder formation.The expression of antiapoptotic gene(Bcl-2) and proapoptotic gene(Bax) was determined by immunohistochemistry.Results At the end of reperfusion,infarct size was reduced by ramipril.TUNEL assay and DNA-laddering study demonstrated that cardiomyocyte apoptosis was attenuated in ramipril-treated hearts,correlating with increased expression of Bcl-2 and ratio of Bcl-2/Bax.Conclusion Ramipril exerts cardioprotection by decreasing the ischemia-reperfusion induced necrosis and apoptosis of cardiomyocyte and increasing the expression of Bcl-2 and ratio of Bcl-2/Bax.

Objective To compare clinical efficacy of clobetasol propionate ointment and vitamin A acid cream in the treatment of skin papule type amyloidosis.Methods 100 cases of skin papule type amyloidosis were randomly divided into the observation group and control group,each had 50 patients.The control group was treated with Vitamin A acid cream while the observation group used clobetasol propionate ointment for treatment.Skin lesion area,infiltration,skin color,skin itching score as well as the cure rate and efficiency were compared after 4 weeks of treatment.Results The two groups' symptoms was improved,symptom scores was decreased gradually after treatment than before,the observation group's symptom scores at 1,2,3,4 weeks were (9.35 ± 1.88),(6.54 ±2.16),(4.08 ±1.32),(2.04 ± 0.95) points which was significant better than (10.86 ± 2.08),(7.98 ± 2.57),(6.25 ± 1.44),(4.56 ± 1.18) points of the control group,the difference of two groups was statistically significant (t =6.22,6.71,7.30,7.41,all P ＜ 0.05) ;4 weeks after treatment,the observation group's efficience and cure rates were 94％ and 34％,significantly higher than 70％ and 22％ of the control group,the difference was statistically significant (x2 =9.040,8.391,all P ＜ 0.05).Conclusion Clobetasol propionate ointment has a exact effect in treatment of skinpapule type amyloidosis than vitamin A acid cream,which is worthy of clinical application.

Objective To explore the effect of hypoxia on exosomes secreted by renal tubular epithelial cells and the function of exosomes in chronic kidney diseases.Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normoxia (21％ O2) or hypoxia(1％ O2) for 48 h was collected and centrifuged gradiently to harvest exosomes.Exosomes were identified and compared by transmission electron microscope, nanoparticle tracking analysis, Western blotting and measurement of the protein concentration.(2) Primary peritoneal macrophages of rats were co-cultured with exosomes in different concentrations (1, 10, 50, 100, 300 mg/L).The expression of interleukin-6(IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) in cells and supernatant were separately detected by quantitative real-time PCR (qRT-PCR) and ELISA, and the expression of phospho (p)-STAT/STAT and suppressors of cytokine signaling 1 (SOCS1) in macrophages was detected by Western blotting.At last, the expression of inflammatory microRNAs (miR) in exosomes was measured by qRT-PCR.Results (1) The vesicles harvested by gradient centrifugation were less than 150 nm and expressed CD63 which was characteristic of exosomes.Hypoxia had no effect on the morphology of exosomes, but stimulated their secretion.(2) Hypoxic exosomes dose-dependently improved the expression of IL-6, TNF-α, iNOS in macrophages polarized by lipopolysaccharide (LPS) and increased the expression of p-STAT while decreased the expression of SOCS1 (P ＜ 0.01).MicroRNAs referred to inflammation such as miR-155 and miR-27a increased in hypoxic exosomes compared to that in normoxic exosomes (P ＜ 0.05).Conclusions Hypoxia makes exosomes promoted the polarization of macrophages to M1, which may account for the microinflammation in chronic kidney diseases.

Aim To investigate the protection of ramipril,BQ-123 and their combination against myocardial ischemia/reperfusion(I/R)injury in vivo in anesthetized rats,and to explore the mechanism of action of drugs on myocardial oxidation-antioxidation system.Methods Healthy male Wistar rats were divided into 5 groups randomly,sham operated(Sham)group,I/R group,ramipril(RAM)group,BQ-123(BQ)group and ramipril and BQ-123(R&B)group.All groups but not sham were subjected to I/R procedure.Twenty four hours before ligation,ramipril(1 mg·kg~(-1))was intragastrically administered to rats in RAM and R&B groups.The equal volume of normal saline was given to rats in other groups.BQ-123(10 μg·kg~(-1)· min-1)was infused intravenously from 10 min before ligation to the end of 30 min ischemia to rats in BQ and R&B groups.The equal volume of normal saline was given to other groups.HR,MAP and the change of ST-segment were observed;ventricular arrhythmias were monitored during ischemia;the infarct size was examined by TTC staining;the activity of myocardial T-SOD,Mn-SOD,CAT and the content of MDA were detected by spectrophotometer.Results Compared with I/R group,the elevation of ST-segment was decreased,onset of VPC and VT was delayed,duration of VPC and VT was shortened,incidence of VPC,VT and VF was decreased,IS and IS/AAR were improved,activity of T-SOD,Mn-SOD and CAT was increased,the content of MDA was decreased in RAM,BQ and R&B groups.Compared with RAM and BQ alone group,onset of VPC and VT,duration of VPC and VT,size,activity of T-SOD and Mn-SOD and content of MDA were changed dramatically in R&B group.Conclusions Ramipril,BQ-123 and the combined use of these two agents protected myocardium from I/R injury in vivo.The protective effects of the combination on delaying onset of VA,shortening duration of VA,decreasing infarct size and content of MDA,and increasing activity of SOD are better than those of using ramipril or BQ-123 alone.

A method for the determination of caffeine in urea was developed based on bubble-in-drop single drop microextraction (BID-SDME) followed by gas chromatography/mass spectrometry (GC-MS).Under the optimum conditions including chloroform as extraction solvent, an exposure volume of 1 μL, a bubble volume of 1.6 μL, stirring for 5 min at 300 r/min, 15% (m/V) NaCl, and a distance of 1 cm between bubble and stirring bar, the detection limit of this method was as low as 0.003 mg/L and the linear range was from 0.005 mg/L to 10 mg/L with correlation coefficient of 0.982.The recoveries of caffeine were from 89.2% to 107.5% at different spiked levels in human urine and the relative standard deviation (RSD, n=6) was less than 8%.

Objective Present study aimed to research the mechanism of the use of 0.2% brimonidine tartrate eye drops in the treatment of optic nerve crush injury of rat.MethodsAnimal models of optic nerve crush injury were created in 60 SD female rats by clipping the exposed optic nerve at 2 mm in retrobulbar for 6 seconds with 78 grams of reverse forceps.The successful model was identified as Marcus-gun pupil without bleeding of fundus after operation.The animals were randomly assigned to model group and brimonidine treating group,and another 30 normal SD rats were used as the normal control group.The 0.2% brimonidine tartrate eye drops was topically administered at 2 hours before operation and after operation twice per day in the brimonidine treating group.The retinas from 18 rats were isolated after 3,7,21 days for RGCs counting by H&E staining,and the retinal ultrastructure was examined under the transmission electron microscope.The retinas from the other 72 SD rats (including normal,model and brimonidine groups) were prepared for the detection of bcl-2 and bax using immunohistochemistry l,3,5,7,14,21 days after the operation.ResultsNormal and almost normal retina structure was exhibited in rats of the normal group and brimonidine treating group,but disorder of cellular arragement and decrease of retinal thickness were found in the model rats under the optical microscope.The RGCs counting was significantly different among the three groups from 3 days through 21 days after operation with the considerably declination in the model group and brimonidine treating group compared with the normal control group (P＜0.05-0.01).However,that of the brimonidine treating group was obviously increased in comparison with the model group (P＜0.01).The expression of bax in rat retina was obviously reduced (P＜0.01),but the expression of bcl-2 was increased in brimonidine group compared with the model group from 5 through 7 days after operation (P＜0.01).ConclusionThe 0.2% brimonidine tartrate eye drops has a preventive effect on optic nerve crush injury of rat,and its inhibition on apoptosis is one of the mechanisms.

BACKGROUND: The existence of minimal residual leukemia cells is the main cause for the recurrence of acute leukemia in children, and immunological biological therapy has attracted more and more attentions in the various methods from eliminating minimal residual disease. Previous studies have found that interferon α-2b can effectively inhibit the increase of tumor cells in vivo in children with neuroblastoma and malignant lymphoma, whether it can inhibit the increase of leukemia cells?OBJECTIVE: To investigate the effects of interferon α-2b in vitro on leukemia cells.DESIGN: A comparative observation taking human promyelocytic leukemia HL-60 cell line as the material.SETTING: Cell Culture Room; Immunological Laboratory; Cell Room, Institute of Pediatrics, Affiliated Hospital,Medical College of Qingdao University.MATERIALS: HL-60 cell line was provided by Shandong Institute of Basic Medical Sciences. Interferon α-2b was purchased from Megagene Company Fluorescein isothiocyanate (FTTC) rabbit-anti-rat Ig solution (CatEK001) and CD13 anti-human monoclonal antibody solution (Cat. DK013Y) were purchased from Union Stem Cell & Gene Engineering Co.,Ltd.METHODS: The experiments were carried out in the Institute of Pediatrics, Affiliated Hospital, Medical College of Qingdao University from March to September 2005. HL-60 cells culture system was established in vitro, and the oncentration was adjusted to 1×109 L-1. The cells were divided into control group and experimental group. In the experimental group, each well was added by interferon-α-2b with the terminal concentration of 5×105, 1×106, 2×106,5×106 and 1×107 U/L, respectively. In the control group, each well was added by saline of the same volume. The cells were cultured continuously for 48 hours. The morphological changes of HL-60 cells were observed using Wright's staining under light microscope; Cell apoptosis was observed using acridine orange/ethidium bromide double staining; Antigen expression and maturation and differentiation on cell membrane were observed by determining CD13 protein expression; Proliferation and activity of HL-60 were detected with methyl-thiazol-tetrazolium (MTT) assay.MAIN OUTCOME MEASURES: The occurrence of apoptosis was judged according to the uniformity and staining of HL-60 nuclear chromatin; HL-60 cell proliferation was judged according to the absorbance (A) value; The maturation of HL-60 cells was judged according to the number of positive CD13 cells.RESULTS: ① HL-60 cell apoptosis: The cells were cultured for 48 hours. When the concentration of interferon α-2b was 5×105 U/L, there were mainly early apoptotic HL-60 cells; When the concentration was 1×107 U/L, there were mainly late apoptotic cells, and the apoptotic rate was significantly higher than those in the control group (P ＜ 0.01 ).② HL-60 cell proliferation: The A values in the experimental groups treated with interferon α-2b of 2×106 U/L and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01). ③ Maturation of HL-60 cells: The percentages of positive CD13 cells in the experimental groups treated with interferon α-2b of 1 ×106 and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01).CONCLUSION: It is concluded that interferon α-2b can enhance the apoptosis, inhibit the proliferation and promote maturation and differentiation of HL-60 cells.

Objective To observe the kinetics of myocardial during isovolumic relaxation (IVR) in normal subjects during exercise stress testing,and evaluate the relationship between the recovery rate of circumferential strain during IVR and the diastolic function of the heart.Methods Echocardiography was performed in 43 healthy students at rest,at peak stress and at recovery of the exercise test.The ratio of peak early-diastolic mitral orifice flow velocity and peak early-diastolic mitral annular velocity in left ventricular posterior wall (E/e) was measured.The records of the movement in the basal,papillary muscle and apical short-axis views of the left ventricular were stored.The circumferential strain during the end systolic (ES) and the end isovolumetric relaxation (IRA) of the subendocardial and subepicardial layers in the three short axis were analyzed using X-strain software.The circumferential strain recovery rate of the subendocardial and subepicardial layers during the IVR (ENCSrec,EPCSrec) were calculated.The circumferential strain of the subendocardial and subepicardial layers in the three short-axis during ES and IRA phases in different status were compared.The relationship between the circumferential strain recovery rate during IVR and E/e were analyzed.Results The circumferential strain of the basal,papillary level in short axis during ES and IRA phases decreases at the beginning and then increased along with the exercise intensity (P ＜0.05).The circumferential strain of the subendocardial layer is greater than the subepicardial layer in the three shortaxis during ES and IRA in different exercise status (P ＜0.01).The ENCSrec and EPCSrec of the three short-axis increases at the beginning and then decreased along with the exercise intensity (P ＜0.05).The ENCSrec and EPCSrec of 3 short axis are correlated with E/e in different exercise phases (P ＜0.05).Conclusions In normal subjects,the kinetics of circumferential strain during IVR is consisted with the physiological changes of the heart during IVR in different exercise phases.The circumferential strains in different myocardial layers are different during IVR.The circumferential strain recovery rate of left ventricular myocardial during IVR may reflex the diastolic function of the heart.

Objective To evaluate the effect of isoflurane on the apoptosis of SH-SYSY cells transfected with APPsw gene and the role of inositol 1,4,5-triphosphate (IP3) recepters.Methods The SH-SYSY ceils transfected with APPsw gene were seeded in culture flasks with the density of 1.2 × 104/cm2.The cells were randomly divided into 4 groups (n =6 each):control group (group C),IP3 receptor antagonist group (group Ⅹ),isoflurane group (group Ⅰ) and isoflurane + IP3 receptor antagonist group (group Ⅰ + Ⅹ).After the cells were cultured for 24 h and attached to the wall,the cells were cultured routinely in group C,and Xestospongin C 100 nmol/L (IP3 receptor antagonist) was added to DMEM culture medium in groups X and Ⅰ + X,and 30 min later the cells were exposed to 1.2 ％ sevoflurane for 8 h in groups Ⅰ and Ⅰ + X.The cells were collected for examination of the ultrastructure and for determination of cell apoptosis,intracellular free calcium ion concentration [Ca2 +] i (by flow cytometry) and expression of IP3 receptor protein (by Western blot).The apoptosis rate was calculated.Results Compared with group C,there was no significant change in the apoptosis rate,[Ca2 +]i or IP3 receptor protein expression in group Ⅹ (P ＞ 0.05),while the cell apoptosis rate and [Ca2 +] i were significantly increased and IP3 receptor protein expression was up-regulated in groups I and Ⅰ + Ⅹ (P ＜ 0.05 or 0.01).Compared with group Ⅰ,cell apoptosis rate and [Ca2+]i were significantly decreased and IP3 receptor protein expression was down-regulated in group Ⅰ + Ⅹ (P ＜ 0.01).The pathological changes of the cells happened in groups Ⅰ and Ⅰ + Ⅹ,and the pathological changes were severer in group Ⅰ than in group Ⅰ + Ⅹ.Conclusion Isoflurane can induce apoptosis of SH-SY5Y cells transfected with APPsw gene through increasing [Ca2+]i and up-regulating IP3 receptor protein expression.