OBJECTIVE : To prepare Piyanjing cream and to establish a quality control method for it. METHODS : Chemical reactions were performed to identify Piyanjing cream qualitatively. The content of main agent, clobetasol propionate was determined by HPLC. RESULTS: Chemical reactions were positive .The clobetasol propionate linearity was between 4.06～97.49?g/ml,and the average recovery was 98.67%(RSD=0.82%,n=9),the average content of clobetasol propionate was 0.492mg/g.CONCLUSION : The prescription and the preparation technique are reasonable,the cream is stable in quality,the quality control is simple ,accurate and practicable.

OBJECTIVE:To determine the contents of the residual solvents including dichloromethane,acetone and ethanol in azithromycin raw material by GC.METHODS:A glass column was used as chromatographic column.The temperature of sample injection was 140℃and column temperature was 160℃.The carrier gas was nitrogen.The column inlet pressure was 30Psi.The detection was performed using hydrogen flame ionization detector.RESULTS:The detection concentrations of dichloromethane,acetone and ethanol were 0.02%～0.1%(r=0.9 995),0.05%～0.25%(r=0.9 994)and 0.02%～0.1%(r=0.9 994),respectively.The average recovery were 99.1%(RSD=1.4%),100.1%(RSD=1.1%)and 99.5%(RSD=1.3%),respectively.All of the three batches of samples were up to the standard with regard to the residual volume of organic solvent residues in azithromycin raw material.CONCLUSION:The method is proved to be accurate,sensitive and reliable,and suitable for the detection of organic solvent residues in azithromycin.

Objective:To establish an HPLC method for determining four constituents ( schaftoside, isoschaftoside, deoxyelephan-topin and 4,5-dicaffeoylquinic acid) in Shennongcha granules. Methods:An HPLC method was performed on a Hypersil C18 column (200 mm × 4. 6 mm,5 μm) with the mobile phase of acetonitrile as the phase A and 0. 025 mol·L-1 phosphoric acid solution as the phase B with gradient elution. The flow rate was 1. 3 ml·min-1 . The detection wavelength was set at 270 nm for schaftoside and isos-chaftoside, 208 nm for deoxyelephantopin and 327 nm for 4, 5-dicaffeoylquinic acid. The column temperature was room temperature. Results:The calibration curve was linear over the concentration range of 5. 850-117. 000 μg·ml-1 for schaftoside, 4. 650-93. 000 μg ·ml-1 for isoschaftoside, 4. 160-83. 200 μg · ml-1 for deoxyelephantopin and 5. 470-109. 400 μg · ml-1 for 4, 5-dicaffeoylquinic acid. The correlation coefficient of all curves was more than 0.999. The average recoverywas 97.70% (RSD=1.40%), 96.87%(RSD=1.13%), 97.53%(RSD =1.69%) and 99.29%(RSD =1.01%) (n =6) , respectively. Conclusion: The developed HPLC method is simple,accurate,and can be used in the content determination of Shennongcha granules.

Objective To observe the expression change of HRG-1 gene between urinary bladder carcinoma and normal tissues,and to investigate the effect of HRG-1-siRNA on the proliferation and apoptosis of human bladder carcinoma cells.Methods Immunohistochemisty was used to detect the expression of HRG-1 in 85 cases of bladder carcinomas and 20 normal bladder tissues.The siRNA of HRG-1 was designed,synthesized,and transfected into bladder cancer cell line T24.Results The HRG-1 gene expression had significant differences between bladder carcinoma and normal bladder tissues (P ＜ 0.05).The positive expression of HRG-1 gene had significant differences between the pathological grades and clinical stages of bladder carcinomas (P ＜0.05).After treated with siRNA,the expression levels of HRG-1 protein and mRNA in T24 cells decreased obviously (P ＜ 0.05).The apoptosis rate of T24 cells transfected with HRG-1-siRNA was significantly different from control-siRNA group and blank group (P ＜ 0.01).Conclusions The high expression of HRG-1 gene may play an important role in bladder carcinoma,and siRNA targeting HRG-1 can suppress HRG-1 protein expression markedly and enhance apoptosis of T24 cells.

Objective To analyze the correlation of perfusion parameters obtained by functional multi-slice computed tomography (fMSCT) with vascular endothelial growth factor (VEGF) detected with immunohistochemistry on rabbit model with VX2 breast cancer and noninvasively evaluate the vascularization of untreated VX2 breast cancer in vivo. Methods Sixteen New Zealand femal rabbits were selected and suspension (1 ml) of mass was injected around the breast areola. CT perfusion was performed after two weeks and perfusion parameters including blood flow (BF), blood volume (BV), mean transit time (MTT)and permeability surface (PS) were assessed. Expression of VEGF in neoplasm was detected with immunohistochemistry. Paried t test was used for the comparison of perfusion parameters between the tumor and muscle and Pearson correlation analysis was used to assess the correlation of VEGF with perfusion parameters. Results The mean value of BF, BV, MTT and PS were (228.21 ± 13. 13 ) ml · min-1100 mg-1 ,(13.45 ± 1.01) ml · 100 mg-1 ,(3. 50 ±0. 20) sand (7.85 ±1.18) ml · min-1 · 100 mg-1 in tumor, respectively. They were (66. 10 ±22. 11 ) ml · min-1 · 100 mg-1 , ( 1.88 ± 1.80) ml · 100 mg-1,(23. 87 ±0. 63)s,(1.55 ±0. 38)ml · min-1 · 100 mg-1 in muscular tissue, respectively. The mean value of BF, BV and PS in tumor were obviously higher than those in muscle, and the mean value of MTT in tumor was lower than that in muscle. There were significant differences in CT perfusion parameters between tumor and muscle (t = 61.83,13.63,27.72,20. 54, P ＜ 0. 01 ). The mean value of VEGF in tumor was 7. 33 ±0. 27 and there were positive correlation with BF ( r = 0. 712, P ＜ 0. 01 ), BV ( r = 0. 647, P ＜ 0. 01 ), PS ( r =0. 627 ,P ＜ 0. 01 ), and negative correlation with MTT ( r = - 0. 564, P ＜ 0. 05 ). Conclusion MSCT perfusion imaging can be used to noninvasively evaluate the vascularization in rabbits with untreated VX2 breast cancer in vivo.

BACKGROUND:Umbilical cord-derived mesenchymal stem cel s are gaining more attention in clinical treatments. Cel viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cel s, it cannot reflect the functional capacity of those cel s accurately because of some subjective factors. OBJECTIVE:To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cel s. METHODS:Human umbilical cord-derived mesenchymal stem cel s were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cel s were preserved in 0.9%saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cel counting kit-8, live-dead assay, cel adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cel s, and the results were compared with colony-forming efficiency, a measure of cel function. RESULTS AND CONCLUSION:Human umbilical cord-derived mesenchymal stem cel s cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cel s were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cel s incubated in the adipogenic and osteogenic medium were both positive. Cel viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cel s measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cel adherent assay-measured viability is the most accurate indicator.

BACKGROUND:The viability of human umbilical cord-derived mesenchymal stem cel s is often declined with the commonly used transplantation storage solution in clinics, which may influence the therapeutic effects of cel ular transplantation. However, reasons for this are stil unknown. OBJECTIVE:To investigate the role of oxidative stress in the reduction of human umbilical cord-derived mesenchymal stem cel s viability in the storage process during clinical transplantation and to observe the effects of radical scavenger on the results. METHODS:Human umbilical cord-derived mesenchymal stem cel s were harvested and cultured in normal saline for 0, 2, 4 and 6 hours at room temperature. Intracel ular reactive oxygen levels were detected at those time points. Antioxidant enzyme activities and levels of malondialdehyde were measured to determine the intracel ular oxidative stress levels after storage. Cel adhesion rate changes were retested after adding N-acetyl cysteine to the storage solution. RESULTS AND CONCLUSION:The reactive oxygen levels in human umbilical cord-derived mesenchymal stem cel s were increased significantly after normal saline storage and levels of malondialdehyde were increased in a time-dependent manner. Activities of superoxide dismutase, catalase and glutathione peroxidase were al reduced. Addition of N-acetyl cysteine into the storage medium decreased the reactive oxygen levels and improved the human umbilical cord-derived mesenchymal stem cel s viabilities. Experimental findings indicate that, increased reactive oxygen species in human umbilical cord-derived mesenchymal stem cel s is one of the reasons for reduced cel viability. Adding the radical scavenger N-acetyl cysteine can improve the storage effects of human umbilical cord-derived mesenchymal stem cel s.

Objective To introduce the technique and evaluate the clinical effect of retroperito-neoscopic anatomical radical nephrectomy. Methods One hundred and sixty-eight patients under-went retroperitoneoscopic anatomical radical nephrectomies. The average tumor size was 4.7 cm (ran-ging from 2.0-6.9 era) in diameter. There were 87 tumors in the left kidney and 81 tumors in right kidney. Ninety-two eases were in cli.nieal stage T1a. N0 M0 and 76 in T1b N0 M0. Retroperitoneal space was created routinely at lateral decubitus position. Four relatively bloodless planes were orderly entered for exposure and separation of the kidney outside Gerota's fascia. The first dissection plane was be-tween the psoas and posterior Gerota's fascia. The renal pedicle was found in this plane. The following dissections proceeded in the plane between posterior Gerota's fascia and fusion fascia. The third dissec-tion plane was between adrenal gland and the upper pole of kidney or between the adrenal gland and di-aphragma. The fourth dissection plane was in the bottom of Gerota's fascia. Results All operations were successfully completed. The mean operative time was 138:J:46 min and estimated blood loss was 90±30 ml. The average day of resuming oral intake was 1.3 d and time of ambulation was 1.2 d. The mean postoperative hospital stay was 5.8 d. Peritoneum injuries happened in 14 patients. Omalgia oc-curred in 18 patients and disappeared 2 d after operation. One hundred and twenty-three patients were followed up, they all survived during the average follow-up of 8 months (ranging from 6-18 months);, .Conclusions Retroperitoneoscopie anatomical radical nephrectomy is a safe and effective procedure. It can decrease operation time, blood loss and complication rate remarkably. It is a good option for patients needing radical nephrectomy.

Objective To investigate the antitumor activities of adenovirus-mediated NDRG2 gene (Ad-NDRG2) and docetaxel on human prostate cancer DU145 cells.Methods The protein expressions of cyclin D1,cycliu E,and NDRG2 in the cells were determined by Western blot.MTT and flow cytometry were used to observe the effects of docetaxel (10-6 mol/L,10-7 mol/L,and 10-s mol/L) and Ad-NDRG2 on prostate cancer cell line DU145 in single or synergistic administration ways for 24 and 48 hours in vitro.Male BALB/C-nu mice with DU145 prostate cancer cell lines were treated by docetaxel and Ad-NDRG2 singly or synergistically in vivo.Results After infected by adenovirus,the protein expression of NDRG2 increased,but cyclin D1 and cyclin E decreased in DU145 cells.Ad-NDRG2 inhibited the cell growth (inhibition ratio =41.8％,t =4.18,P ＜0.01),promoted apoptosis (apoptosis ratio =32.4％,x2 =11.66,P ＜0.05),changed the ratio of G2/M phase from 50.2％ to 23.6％,and reversed partially the G2/M arrest,of DU145 cells induced by 10-7 mol/L docetaxel.In vivo experiment showed that docetaxel,Ad-NDRG2,and combination of docetaxel and Ad-NDRG2 inhibited tumor growth with a inhibition rate of 30.7％,28.2％,and 55.8％,respectively.The coefficient of drug interaction (CDI) of docetaxel and Ad-NDRG2 was 0.89.Conclusions Ad-NDRG2 can enhance the growth suppression and apoptosis induced by docetaxel in synergistic way in vitro and in vivo.It demonstrated the great potential of Ad-NDRG2 in the treatment of androgen-independent prostate carcinoma.

Objective To determine the value of applying LIIR Automatic Analysis System of Infrared Spectroscopy in analyzing urinary stone composition. Methods 1450 samples of urinary stones were collected from 1032 male and 418 female patients. The age of patients ranged from 6 months to 88 years. The mean ages were 41.7±15.3 and 42.0±15.6 years for male and female patients, respectively. Of 1450 stones, 875 cases were located in kidney (60.34%), 504 cases in ureter (34.76%) and 71 cases in bladder (4.90%). All stones were analyzed by LIIR Automatic Analysis System of Infrared Spectroscopy (Tianjin). Analysis results were reevaluated by the artificial analysis of spectrogram, if necessary, with polarization microscope, chemical analysis, and X-ray diffraction.Results Calcium oxalate monohydrate stones were found in 714 cases (49. 24%), carbonate apatite stones in 444 cases (30.62%), anhydrous uric acid stones in 93 cases (6.41%), calcium oxalate dihydrate stones in 92 cases (6. 34 % ), ammonium magnesium phosphate hexahydrate stones in 28 cases (1.93%), cystine stones in 23 cases (1.59%), ammonium urate stones in 20 cases (1.38%), uric acid dihydrate stones in 16 cases (1.10%), brushite stones in 12 cases (0.83%), sodium urate monohydrate stones in 2 cases (0. 14%), calcium carbonate stones in 1 cases (0. 07%), and other stone types in 5 cases (0. 34%). Most urinary stones were composed of 2 or more compositions, and pure stones were only observed in 397 cases (27.38%). Most of the mixed stones contained calcium and non-calcium mixed stone was rarely observed. In addition, 15 stones were found in infants who had consumed melamine-contaminated milk powder. These stones were composed of uric acid dihydrate and ammonium urate. The results of reevaluation by artificial analysis showed the following: among pure and mixed stones, false detection occurred in 6 cases (0.41%), of which the composition was ammonium urate or carbonate apatite determined by automatic system but the true composition was anhydrous uric acid. False negative detection occurred in 9 cases (0.62%), of which the composition was ammonium magnesium phosphate hexahydrate or carbonate apatite in 7 cases, but in other 2 cases the composition could not be determined by artificial analysis. The false negative detection of components with relatively low content occurred in 6 cases and 10 cases in stones with 2 components and 3 components, respectively. The undetected composition in these cases was ammonium magnesium phosphate hexahydrate or carbonate apatite. Conclusion Automatic Analysis System of Infrared Spectroscopy has many advantages in accuracy, automation and is quick in analyzing the composition of urinary stones, and is worthy of promotion in clinical use.