Objective To explore the inhibitory effect of astragalus polysaccharide (APS) on the proliferation of human erythroleukemia K562 cells and its mechanisms.Methods After K562 cells (purchased from Shanghai cell bank of chinese academy of science) were treated with different concentrations (0 mg/L,100 mg/L,200 mg/L and 400 mg/L) of APS.The influences of APS on the growth rate,doubling time and cell cycle distribution of K562 cells were observed by methyl thiazolyl tetra-zolium assay (MTF) and flow cytometry,respectively.Furthermore,the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting assay were used to detect the expressions of Cyclin A,Cyclin B,Cyclin E and p21 gene at the mRNA and protein levels,respectively.Results MTT assay findings showed that,compared to the control group (0 mg/L APS),growth rates of K562 cells treated with 100 mg/L,200 mg/L and 400 mg/L APS decreased significantly (all P ＜ 0.01),and the doubling times lengthened significantly (all P ＜ 0.01).Flow cytometry findings revealed that,compared to the control group,the G1 phase cells in K562 cells of APS group increased significantly (P ＜0.01),while the S and G2/M phase cells decreased significantly (all P ＜ 0.01).RT-PCR and Western blotting results indicated that Cyclin B and Cyclin E expression of K562 cells at the mRNA and protein levels in the APS group were significantly lower than those of the control group(all P ＜ 0.01),whereas p21 expression was significantly enhanced at mRNA and protein levels (P ＜ 0.01),and Cyclin A expression was not significantly different at mRNA and protein levels between the 2 groups (all P ＞ 0.05).Conclusions APS could inhibit the proliferation of human erythroleukemia K562 cells.APS could inhibit the proliferation of K562 cells by down-regulating the expression of Cyclin B and Cyclin E and up-regulating the expression of p21.

Adult ; Chemotherapy, Adjuvant ; Diagnosis, Differential ; Female ; Humans ; Lung Neoplasms ; secondary ; Mediastinal Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; surgery ; Mediastinum ; diagnostic imaging ; pathology ; Neoplasm Recurrence, Local ; Osteosarcoma ; diagnostic imaging ; drug therapy ; pathology ; secondary ; surgery ; Tomography, X-Ray Computed

Objective: To observe the clinical efficacy of heat-sensitive moxibustion for vascular dementia and explore its mechanism. Methods: A total of 70 patients with vascular dementia were randomized into an observation group and a control group, with 35 cases in each group. The control group was treated with piracetam, and the observation group was treated with heat-sensitive moxibustion on the basis of the treatment of the control group. The treatment lasted for 8 weeks. Before and after the treatment, the mini-mental state examination (MMSE), activity of daily living (ADL) and traditional Chinese medicine (TCM) symptom scores were assessed, and the levels of acetyl choline (Ach) and homocysteine (Hcy) were measured. The efficacy was evaluated after treatment. Results: The total effective rate of the observation group was significantly higher than that of the control group (P<0.05). After treatment, the MMSE and ADL scores in the observation group decreased significantly, and were lower than those in the control group (all P<0.05); the TCM symptom score of the observation group decreased significantly (P<0.05), while that of the control group had no significant change (P>0.05); the plasma Ach level in the observation group increased significantly (P<0.05), and the Hcy level decreased significantly (P<0.05), which were statistically different from those in the control group (both P<0.05). Conclusion: Heat-sensitive moxibustion plus piracetam is effective in treating vascular dementia. It can significantly improve dementia symptoms and ADL, which may be related to the correction of plasma Ach and Hcy levels.

Objective: To observe the clinical efficacy of heat-sensitive moxibustion in intervening acute ischemic stroke. Methods: A total of 112 patients were divided into a control group and an observation group by the random number table method, with 56 cases in each group. The control group was treated with conventional treatment, and the observation group was treated with heat-sensitive moxibustion on the basis of the treatment in the control group. The National Institute of Health stroke scale (NIHSS) and modified Barthel index (MBI) were evaluated, and the intracranial hemodynamic indicators including mean velocity (Vm), pulsatility index (PI) and resistance index (RI) were recorded, and the serum levels of superoxide dismutase (SOD) and homocysteine (HCY) were measured before and after treatment. Clinical efficacy was evaluated after treatment. Results: The total effective rate of the observation group was significantly higher than that of the control group (P<0.05). After treatment, the NIHSS scores in both groups decreased significantly (both P<0.05), and the MBI scores increased significantly (both P<0.05). The improvements of NIHSS and MBI scores in the observation group were superior to those in the control group (both P<0.05). Vm in both groups increased significantly (both P<0.05), PI and RI decreased (all P<0.05), and Vm, PI and RI in the observation group were superior to those in the control group (all P<0.05). The serum levels of SOD and HCY in the observation group were significantly improved, and were statistically different from those in the control group (both P<0.05). Conclusion: Conventional treatment plus heat-sensitive moxibustion is effective in intervening acute ischemic stroke. It can promote the recovery of neurological function, improve daily activities, and improve intracranial blood flow, which may be related to the regulation of serum SOD and HCY levels.

Objective: To study the quality and transdermal properties of matrine microemulsion-based hydrogel (MBH) to provide basis for the development of the preparation.Methods: The stability of MBH was observed at 4 ℃ for 3 months and the changes of particle appearance, viscosity, pH and matrine content were observed.The transdermal permeation of MBH was investigated by a dual chamber permeation and diffusion device with excised mouse skin as the barrier.Taking rabbits as the experimental subjects, the irritation of MBH to the normal skin and damaged skin was investigated.Results: The appearance, viscosity, pH and matrine content of MBH at 4 ℃ in 3 months did not change significantly.In vitro transdermal test showed that MBH had a good penetration rate on mouse skin, and no skin irritation occurred after single or multiple administrations.Conclusion: MBH has good stability and high rate of transdermal penetration without skin irritation, which is a promising drug delivery system of matrine with good application prospects.

AIMTo investigate the cellular uptake of monostearin solid lipid nanoparticles (MSLN) and the influence on the cellular uptake by MSLN modified with PEG2000 in human-type II cell alveolar epithelial cell line (A549) and murine macrophages cell line (J774A1).

METHODSMSLN were prepared by a novel solvent diffusion method. The particle size distribution and zeta potential of MSLN, measured by light scattering and electrophoretic mobility, were investigated. The cytotoxicity of MSLN and MTX-loaded MSLN in A549 cells were performed by the MTT method. Rhodamine B was incorporated into solid lipid nanoparticles as fluorescent marker, after PEG2000 integrating monostearin during preparation, the cellular uptake of MSLN by A549 and J774A1 cell lines were determined spectrofluorimetrically.

RESULTSThe IC50 of MSLN and MTX-loaded MSLN on A549 cells were 227.56 microg x mL(-1) and 71.37 microg x mL(-1), respectively. The percentage of cellular uptake showed a negative correlation to the concentration of MSLN in incubation medium and the internalization behaved rapidly. Contrary to situation in J774A1 cell line, internalization of solid lipid nanoparticles was promoted with increasing the content of PEG2000 incorporated into MSLN in A549 cell line.

CONCLUSIONAfter modifying MSLN with PEG2000, it represents relative lower phagocytosis by J774A1 cell line and higher uptake in A549 cell line.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Cell Line ; Cell Line, Tumor ; Cell Survival ; drug effects ; Drug Carriers ; Drug Delivery Systems ; Glycerides ; chemistry ; metabolism ; toxicity ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Macrophages ; cytology ; Mice ; Nanotechnology ; Particle Size ; Phagocytosis

Adult ; Antibodies, Monoclonal ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Follow-Up Studies ; Humans ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; Male ; Mucin-1 ; metabolism ; Prednisone ; therapeutic use ; Receptor Protein-Tyrosine Kinases ; metabolism ; Vincristine ; therapeutic use

Objective To observe effect of lidocaine pretreatment to malondialdehyde(MDA) and endothelin(ET) of patient ac-cepeted brain tumor removing and discuss the optimized pretreatment time .Methods 60 brain tumor patients in the hospital from March 2009 to September 2011 .according to the different pretreatment time ,the patients were randomly divided into five groups :group A(preoperative 48 h) ,group B(preoperative 24 h) ,group C(preoperative 12 h) ,group D(0 h or anesthesia induced) ,group E (control group) and group F(blank control group) ,10 cases in each group .Group A ,B ,C ,D with 1% lidocaine 1 .5 mg/kg intrave-nous pretreatment on schedule ,then induced conventional anesthesia ;group E were supplemented with 1% lidocaine 2 .5 mg · kg -1 · h-1 intravenous injection after anesthesia induction ;group F was performed routine program without lidocaine .The spontaneous breathing time ,awake time and tracheal extubation time was recorded ,while NIHSS score for evaluation of neural function defect was applied ,and peripheral serum level of MDA and ET was detected by colorimetric technique and radio-immunity .Results In group C ,the spontaneous breathing time ,awake time and tracheal extubation time were shorter than other groups ,but the difference had no statistically significant(P>0 .05) .There was no significant difference among each group in the aspect of NIHSS score 1 day before surgery(P>0 .05) ,after 14 days of operation ,NIHSS of group C was statistically lower than that of group E and group F (P<0 .05) .Before anesthesia induction ,there was no significant difference among groups (P> 0 .05) .MDA and ET content in group C was significantly lower than those in other groups after surgery (P<0 .05) .Conclusion Lidocaine given 12 h before cere-bral ischemia has varying degree protection against cerebral ischemia-reperfusion injury .The protection has relation with the de-crease of MDA and ET content .

The enantiomers separation of thirteen drugs collected in Ch.P2010 was performed on chiral stationary phase of cellulose ramification (chiralpak OD and chiralpak OJ) by high performance liquid chromatographic (HPLC) methods,which included ibuprofen (Cl),ketoprofen (C2),nitrendipine (C3),nimodipine (C4),felodipine (C5),omeprazole (C6),praziquantel (C7),propranolol hydrochloride (C8),atenolol (C9),sulpiride (C10),clenbuterol hydrochloride (C11),verapamil hydrochloride (C12),and chlorphenamine maleate (C13).The mobile phase consisted of isopropanol and n-hexane.The detection wavelength was set at 254 nm and the flow rate was 0.7 mL/min.The enantiomers separation of these thirteen racemates on chiralpak OD column and chiralpak OJ column was studied,while the effects of proportion of organic additives,alcohol displacer and temperature on the separation were studied.And the mechanism of some of racemates was discussed.The results indicated that thirteen chiral drugs could be separated on chiral stationary phase of cellulose ramification in normal phase chromatographic system.The chromatographic retention and resolution of enantiomers could be adjusted by factors including column temperature and the concentration of alcohol displacer and organic alkaline modifier in mobile phase.It was shown that the resolution was improved with reducing concentration of alcohol displacer.When concentration of organic alkaline modifier was 0.2％ (v/v),the resolution and the peak shape were fairly good.Most racemates mentioned above had better resolution at column temperature of 25 ℃.When racemates were separated,the temperature should be kept so as to obtain stable separation results.

Objective To evaluate the selection and outcome of procedures for the treatment of female patients with low urovaginal fistulas.Methods Between Jan.1999 and Dec.2012,a total of 94 low urovaginal fistula patients with mean age 28(5-58)years and the duration of the condition for mean 4(1-23)years were treated using a variety of procedures.Of the 94 patients,the etiology was trauma in 57 patients,iatrogenic injuries in 34,local inflammation in 2 and congenital in 1.Urethral stricture was associated with urethrovaginal fistulas in 61 patients(Group of urethra)and vesicovaginal fistula in 33 (Group of bladder).Of the group of urethra,it was associated with ileovaginal fistula in 2 patients,rectovaginal fistula in 3 and veginal strictures in 8.Of the group of bladder,the fistula was simple or incipient in 15 cases and complex or recurrent in 18 cases.In group of urethra,pedicle labial skin grafs urethroplasty was used in 30 cases,island flap of vulva urethroplasty in 4 cases,vaginal wall flap urethroplasty in 18 cases,end to end anastomotic urethroplasty in 6 cases,and anterior bladder flap uretbroplasty in 3 cases.At the same time of urethroplasty,bladder neck reconstruction was performed in 6 cases with pre-existing traumatic sphincter incompetence,intestovaginal fistula repair was performed in 5 cases.In the group of vesicovaginal fistula,the fistula repairing was performed by transabdominal approach in 18 cases and by transvaginal approach in 15 cases.Results There were no serious complications postoperatively.Patients were followed up with mean 45(5-140)months.Of the 34 cases underwent pedicle labial or an island flap of vulva urethroplasty,fistulas was recurrent in 3 cases and urethral strictures was happened in one case,3 cases had frequent and stress incontinence,however,all 3 cases achieved urinary continence in 3-8 months postoperatively;of the 18 cases underwent urethroplasty using vaginal flap repairs,all patients achieved anatomical success,and continence in 15 patients and stress incontinence in 3 patients;of the 6 patients underwent urethral end to end anastomosis,all patients achieved urethral anatomical repair success and the continence was achieved in 5 patients,stress incontinence in one patient;and the left 3 patients underwent bladder anterior wall urethroplasty,urethral anatomical repair success was in all 3 patients and continence in one,stress incontinence in one and dysuria in one.Endoscopic resection was performed in the patient with voiding difficulty,after which the patient could void smoothly.In the group of urethra,successful urethral anatomical recovery rate was 91.8％(56/61)and successful functional recovery rate was 83.6％(51/61).In the group of bladder,the fistulas were successfully repaired by one procedure in 29 patients(87.9％)and recurrence in 4 patients.Conclusions The selection of procedures for treatment of female patients with low urovaginal fistulas should be determined by stricture characteristics,including location,length and vaginal condition.It is very important to prevented fistulas reformation during operation by using tissue flaps as a bulk.