OBJECTIVE:To improve the HPLC method for the content determination of Methotrexate(MTX)for injection.METHODS:The determination was performed on Inertsil C 8 column,mobile phase consisted of acetonitrile-water-acetic acid glacial-triethylamine(14:86:0.5:0.3)at flow rate of 0.8 mL?min-1.UV detection wavelength was set at 306 nm.Metronidazolum was selected as internal standard for internal standard method to replace external standard method stated in Chinese Pharmacopeia.Result of internal standard method was compared with that of external standard method.RESULTS:The linear range of MTX was 0.52～3.64?g?mL-1(r=0.999 7)with an average recovery of 99.77%(RSD=0.42%).Two kinds of methods obtained similar results.CONCLUSION:Improved method is convenient,accurate and reproducible for quality control of MTX.

OBJECTIVE To explore the rational application of aminoglycosides antibiotics.METHODS We randomly selected 520 patients' records of applying aminoglycosides antibiotics from Jan to Dec in 2007.Types of antibiotics,dosages,usage,ways of delivery and periods and combination of medicines were analyzed.RESULTS Out of the 520 cases,the oldest patient was 89 year and the youngest one was 7 years.The application rate of amikacin was the highest(35.6%),which was delivered mainly by iv,the combination of medicines accounted for 92.7%.CONCLUSIONS The application of aminoglycosides antibiotics in our hospital is basically rational.In some cases the improper antibiotics are used and the duration is a little longer,which should be supervised.

OBJECTIVE:To establish an ion-pair RP-HPLC method for determination of the concentration of netilmicin sulfate in human serum.METHODS:The determination was performed on Luna C18 column with column temperature set at 40℃;the mobile phase consisted of acetonitrile-water (15∶85,pH 2.0,containing 10 mmol?L-1 sodium heptanesulfonate) at a flow rate of 0.8 mL?min-1.The detection wavelength was set at 205 nm.RESULTS:The linear range of netilmicin was 2～20 mg?L-1(r=0.997 6) with a recovery rate of 93.66%～96.05%.The intra-day and inter-day RSD were both less than 10%,respectively.CONCLUSION:The method is sensitive and simple with accurate and reliable results,and it is able to satisfy the requirement for the determination of serum concentration of netilmicin.

Objective To evaluate the soothing effects of hemodiafiltration on patients with refractoriness nephrotic edema and its influence on prognosis of nephrotic syndrome.Methods All the 15 cases involved had undergone hemodiafiltration periodically during acme edema and administered with the same standard loading dose of prednisone for 8 weeks.Meanwhile,clinical and lab indexes (urine volume,Cr,Bun,urinary protein,albumin) were measured.Results All patients improved in urine volume and renal function gradually after the treatment of hemodiafiltration for 1 to 3 times.By the 8th week of post-hemodiafiltration,urinary protein 24 h declines obviously[(6.42?2.31)g/d to (0.87?1.24)g/d].Conclusion Hemodiafiltration is an effective treatment in relieving retention of water,increasing the remission rate of protein urine,recovering patients' renal function and improving their prognosis to nephritic syndrome patients with severe refractory edema.

Objective To evaluate septin-9 and clusterin protein levels in the peripheral blood samples from epithelial ovarian cancer patients, and explore its clinical significance. Methods Clinical data of 200 patients in Cancer Hospital,Chinese Academy of Medical Sciences from Jan. 29, 2008 to Feb. 1,2010 were collected. The peripheral blood samples were obtained from 137 epithelial ovarian cancer patients, 12 borderline ovarian tumor patients, 10 benign ovarian tumor patients, 41 benign pelvic lesion patients and 58 healthy women. The septin-9 and clusterin protein levels in the plasma were measured by double antibody sandwich ELISA or ELISA. The clinical significance of clusterin and septin-9 in plasma was analyzed. The diagnostic efficacy of septin-9 and clusterin protein in the detection of ovarian cancer was evaluated by the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Results Double antibody sandwich ELISA showed: the mean levels of plasma septin-9 in epithelial ovarian cancer patients or benign pelvic lesion patients were significantly higher than that in healthy women detedted by double antibody sandwich ELISA (P<0.01). The mean levels of plasma septin-9 in epithelial ovarian carcinoma patients with tumor family history or distance metastasis were significantly higher than those patients without (P<0.05). While the expression level of septin-9 protein in peripheral blood of ovarian cancer patients was not related to the patient age, pathologic stage, pathologic differentiation, smoking history, treatment history (including surgery, radiotherapy and chemotherapy) and lymph node metastasis (all P>0.05). ELISA showed: the mean level of plasma clusterin in epithelial ovarian cancer patients was significantly higher than that in healthy women deteded by ELISA (P=0.021). The expression level of clusterin protein in peripheral blood of ovarian cancer patients was not related to the above clinical pathological parameters (all P>0.05). To distinguish between ovarian cancer patients and healthy women by septin-9 protein expression level in plasma, when AUC was 0.712 and cut off was 0.28, the sensitivity of detection ovarian cancer by septin-9 protein expression was 82.5%, and the specificity was 50.0%. To distinguish between ovarian cancer patients and healthy women by clusterin protein expression level in plasma, when AUC was 0.636 and cut off was 87.96 pg/L, the sensitivity of detection ovarian cancer by clusterin protein expression was 71.5%, and the specificity was 41.4%. Conclusions The expression of septin-9 and clusterin protein in peripheral blood of ovarian cancer patients is increased, especially the expression level of septin-9 protein with related to the distant metastasis. The study results shown that the detection of septin-9 and clusterin in plasma has a certain diagnosis value in ovarian cancer, which may be a potential markers for ovarian cancer.

Objective To investigate the clinical features, treatments and prognostic factors of rhabdomyosarcoma (RMS) in the female genital tract. Methods A retrospective analysis was performed on 13 cases of RMS in the female genital tract. Clinical characteristics, treatments and prognosis were compared and analyzed statistically. Results The median age was 21.0 years (range, 6 to 54 years). There were 6 cases vaginal RMS and 7 cases cervical RMS, included 11 cases of embryonal RMS (ERMS) and 2 cases of alveolar RMS (ARMS). According to the Federation International of Gynecology and Obstetrics (FIGO)staging system,there were 6 cases of stageⅠ, 3 cases of stageⅡ, 1 case of stageⅢand 3 cases of stage Ⅳ, the median survival time were respectively 112.5, 153.0, 9.0 and 3.5 months. According to the Intergroup Rhabdomyosarcom Study Group (IRSG) staging system, there were 10 cases of stageⅠ and 3 cases of the stage Ⅳ, and their median survival time were respectively 112.5 and 3.5 months. Nine patients received surgery and the median survival time was 108.0 months (range, 9 to 228 months), 6 of them received chemotherapy after surgery and the median survival time was 152.0 months (range, 9 to 228 months), the other 3 cases did not receive any therapy after surgery and the median survival time was 25.0 months (range, 9 to 108 months). Four patients did not receive surgery and the median survival time was 6.3 months (range, 1 to 117 months). There were 2 cases received combined treatment included radiotherapy and the survival time were respectively 4 and 198 months. There were 8 cases who was survival without disease and 5 cases died of cancer. The median survival time in 13 patients was 25.0 months (range, 1 to 228 months) and the 5-year overall survival rate was 58.6%. Conclusions The prognosis of early stage of RMS in the female genital tract is good. While, the prognosis of advanced stage is poor. The standard treatment strategy is combination of surgery and chemotherapy,whether radiotherapy could improve the prognosis still need further study.

OBJECTIVE:To prepare Xielong liquor and establish its quality control.METHODS:Xielong liquor was prepared by classic merceration.Paeonia lactiflora in the formulation was identified by TLC,and the content of the active component ferulaic acid was determined by HPLC.RESULTS:The preparation was yellowish-brown in color and its TLC spots were clear and well-isolated.The linear range of ferulaic acid was 3.66～36.64 mg?L-1(r=0.999 8)and its the average recovery was 100.47%(RSD=0.84%,n=15).CONCLUSION:The preparation technology of Xielong liquor is simple and feasible and the quality of the preparation is stable and controllable,thus the established standard is applicable for the quality control of Xielong Liquor.

Objective:To study the effects of tamoxifen on proliferation and ER expression of human hepatocellular carcinoma cells.Methods:HepG2 cells were treated with tamoxifen at different concentration and different action time.MTT was used to determine the suppression rate of human hepatocellular carcinoma cell.The effects of tamoxifen on human hepatocellular carcinoma cell ER performance were observed by immunohistochemistry.Results:Tamoxifen inhibited the proliferation of human hepatocellular carcinoma cells and suppressed human hepatocellular carcinoma cell ER performence.Conclusions:Tamoxifen may suppress human hepatocellular carcinoma cell proliferation ER performance.

Objective To explore the effects of vitamin C and niacinamide on the growth and differentiation of human primary cultured keratinocytes.Methods Normal human foreskin was used in this study.The epidermis was separated enzymatically from the dermis by thermolysin,and keratinocytes were isolated from the epidermis by digestion with trypsin plus EDTA.The single keratinocytes were cultured with undedying NIH-3T3 cells as feeder cells in a complete medium supplied with 50 mg/L (vitamin C group),niacinamide of 400 μmol/L(niacinamide group)or vehicle(control group).Immunocytochemistry and immunodot blot were performed using monoclonal antibodies directed against C-myc,cyclin D1,filaggrin and involucrin.Results The colony number was highest in vitamin C group,followed by the control group and niacinamide group,and the colony morphology in vitamin C group was similar to that in the control group,but distinct from that in the niacinamide group.A significant increase was noticed in the expression of C-myc,cyclin D1,filaggrin and involucrin in vitamin C-treated keratinocytes compared with the control keratinocytes(all P＜0.05);however,in niacinamide-treated keratinocytes,the expression of filaggrin was significantly enhanced(P＜0.01),that of involucrin remained unchanged(P＞0.05),while that of C-myc was depressed(P＜0.05).Conclusions These results demonstrate that vitamin C has a favorable effect on both the growth and differentiation of human keratinocytes,while niacinamide seems to only promote the differentiation but attenuate the growth of human keratinocytes.

Objective To explore the value of gemstone spectral imaging (GSI) in the detection and differential diagnosis of benign and malignant thyroid nodules.Methods The spectral CT images in 88 patients with thyroid nodules were analyzed retrospectively.All the patients underwent spectral CT scanning,including 33 patients undergoing biphase enhanced CT imaging.Nodules were divided into benign and malignant ones according to histopathologic results.The iodine concentration,slope of spectral curve,effective atomic number in non-enhanced and enhanced scanning were compared between benign and malignant group by the Wilcoxon rank sum test,respectively.The optimal iodine concentration threshold to predict malignancy was obtained by receiver operating characteristic curve (ROC),sensitivity and specificity were achieved.Results A total of 106 nodules were detected,including 76 benign and 30 malignant nodules.In non-contrast CT imaging,the average iodine concentration,slope of spectral curve,effective atomic number of were 2.35 × 100 μg/ml,0.29 and 7.71 for benign group:-0.51 × 100 μg/ml,-0.06 and 7.52 for malignant group (Z value were-3.072,-3.107 and-3.055,respectively ; P ＜ 0.05).In the arterial phase,the average iodine concentration,slope of spectral curve,effective atomic number of the two group were 27.22 × 100 μg/ml,3.23 and 9.10 for benign group; 18.81 × 100 μg/ml,2.24 and 8.69 for malignant group (Z value were-2.582,-2.582 and-2.564,respectively; P ＜ 0.05).In venous phase,no significant difference was found for each parameter between the two groups (P ＞ 0.05).The optimal iodine concentration to predict malignancy was-0.35 × 100 μg/ml in non-enhanced phase with 56.7％ sensitivity and 73.7％ specificity.The optimal iodine concentration was 22.91 × 100 μg/ml in arterial phase,with 76.2％ sensitivity and 75.0％ specificity.Using iodine concentration to predict malignancy in both noncontrast phase and arterial phase,the sensitivity and specificity were 81.2％ and 73.3％,while the accuracy was 81.6％.Conclusion Gemstone spectral CT imaging can quantitatively evaluate the iodine content of thyroid nodules,having a potential value in differential diagnosis of thyroid nodules.