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 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 effects of natural avocado oil and olive oil on the proliferation and differentiation of HaCaT cells. Methods MTT assay was performed to determine the optimal work concentration of avocado oil and olive oil. Cultured HaCaT cells were divided into 3 groups, i.e., avocado oil group treated with avocado oil of 3% (v/v), olive oil group treated with olive oil of 3% (v/v), and control group without any treatment. Immunocytochemistry and immuno-dot-blot method were used to detect the expressions of c-myc, mitogen-activated protein kinase ( MAPK ), nuclear factor ( NF)-κB, filaggrin, involucrin and keratin10 in HaCaT cells. Results As immunocytochemistry showed, the mean grey values (staining intensity) of c-myc,MAPK, and NF-κB in HaCaT cells were 131.4 ± 6.6,136.3 ± 4.5 and 134.3 ± 5.2 respectively in the avocado oil group, 121.1 ± 4.5, 107.9 ± 7.3 and 106.4 ± 5.4 respectively in the olive oil group, significantly higher than that in the control group (101.9 ± 8.9,91.4 ± 5.1 and 94.3 ± 7.0, respectively, all P＜ 0.05), and the avocado oil group was higher than the olive oil group in all the above parameters (all P ＜ 0.05). Increased expressions of filaggrin, involucrin and keratin 10 were observed in the avocado oil group and olive oil group compared with the control group (all P＜ 0.05), and in the olive oil group than in the avocado oil group (all P＜ 0.05).The mean grey values of these proteins obtained by immunocytochemisty were significantly correlated with those obtained by immuno-dot-blot method in avocado oil group (r = 0.94, P ＜ 0.01 ) and olive oil group (r=0.97, P ＜ 0.01 ). Conclusions Certain concentrations of avocado oil and olive oil can promote the proliferation and differentiation of HaCaT cells; avocado oil is more capable to accelerate their growth and proliferation, and olive oil to enhance their differentiation.

Objective To discuss the clinical characteristics,treatment and prognostic factors of brain metastasis from gynecological cancers.Methods Clinical records of 25 brain metastasis patients from gynecological cancers admitted from January 1999 to January 2009 were reviewed retrospectively.The curative effects of different treatments were compared.The prognostic factors were determined using the Cox regression model.Results There were 14 cases ovarian malignant tumor,6 cases cervical carcinoma and 5 cases uterine malignant tumor.Seven cases (28％,7/25 ) had solitary metastatic lesion in the brain.Extracranial metastases were detected in 18 cases (72％,18/25 ).Five patients only received one kind of treatment,and the mean survival time was 4.0 months (0.5 to 9.5 months).Sixteen patients received combined treatment,3 of them received combined treatment including surgery,and the mean survival time was 8.4 months (4.1 to 13.4 months) ; 13 of them received chemotherapy and radiotherapy,and the mean survival time was 14.2 months (4.3 to 58.1 months).Four patients received only palliative supportive care,and the survival time was only 0.1 to 1.0 month.The Cox regression model showed that Karnofsky performance status scale,with or without extracranial metastases and the treatment method were the independent prognostic factors of brain metastasis from gynecological cancers ( P ＜ 0.05 ).Conclusion The incidence of brain metastasis from gynecological cancers is low and the prognosis is very poor.Combined treatments may get better effects.

Objective To discuss the diagnostic value of monochromatic energy image spectral CT in active small bowel bleeding and to screen the optimal energy level that indicates active bleeding of the gastrointestinal tract.Methods The bleeding model was established using the small intestines of pigs in vitro.Seven blood flow rates were simulated:0.500,0.400,0.300,0.200,0.100,0.050,0.025 ml/min,respectively.For each rate,a GE Discovery HD750 CT scanner was used in GSI scan mode and 64 slice CT was performed,with a delay of 15 s and 40 s simulated the arterial phase and portal venous phase,respectively.Each out of the blood flow rate in the 2 modes was respectively scanned 5 times.The GSI reconstruction platform was employed to obtain 7 monochromatic energy images(40,50,60,70,80,90,100 keV).A set of polychromatic energy images was obtained from an ordinary scan.The detection rates of the contrast agent exudation regions using the two scanning methods were compared.The contrast to noise ratios(CNR) for the contrast agent exudation regions were measured.Randomized block analysis of Variance was performed to compare the differences in CNR between energy levels.The x2 test was used to compare the detection rates obtained from the 2 scanning methods.Results The detection rates for energy spectral CT and 64 slice CT in the arterial phase were 31/35 and 23/35,respectively; there was significant difference(x2=5.185,P=0.023).The total detection rates of portal venous phase were 35/35 and 32/35,respectively,there was no significant difference(x2=l.393,P=0.238).On the ordinary scan mode,the detection rates of arterial and portal venous phase difference was statistically significant(x2 =6.873,P =0.009);but on the GSI scan mode,there was no significant difference(x2=2.386,P=0.122). The CNR values at 8 group energy levels for arterial phase and portal venous phase were statistically different(respectively P＜ 0.05),the CNR value of the contrast agent exudation regions at 50 keV and 60 keV monochromatic energy images on the GSI scan mode higher than that of polychromatic energy images,the difference was statistically significant(respectively P＜0.05),the CNR in portal venous phase images were higher than that of arterial at all energy groups,differences were statistically significant(t=-3.996 to-2.380,respectively P＜ 0.05).Conclusions Monochromatic energy image spectral CT demonstrates superiority over polychromatic energy images in detecting active bleeding of the gastrointestinal tract.The optimal monochromatic energy value for detection was between 50 keV and 60 keV,and the detection was easier in the portal venous phase than in the arterial phase.

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.

BACKGROUND:Liquid chip techniques (Luminex) is a recently rising method for detecting anti-panel reactive antibody, which is characterized by high sensitivity, and strong specificity, less interference and high flux.
OBJECTIVE:To compare the sensitivity and detection difference of panel reactive antibody in serum of kidney disease patients detected by enzyme-linked immunosorbent assay and Luminex.
METHODS:Serum samples of 280 patients with kidney disease were selected. The enzyme-linked immunosorbent assay and Luminex methods were used to measure positive rate of panel reactive antibody. Chi-square test for fourfold table data was utilized for statistical analysis.
RESULTS AND CONCLUSION:The positive rates of panel reactive antibody were respectively 18.9%and 33.6%as detected by enzyme-linked immunosorbent assay and Luminex method. The positive rates of anti-HLA-I antibody and anti-HLA-II antibody were respectively 12.8%and 12.5%, as detected by enzyme-linked immunosorbent assay. The positive rates of anti-HLA-I antibody and anti-HLA-II antibody were respectively 25.0%and 20.7%, as detected by Luminex method. Positive detection rate was significantly higher in the Luminex group than that in the enzyme-linked immunosorbent assay group. Moreover, Luminex method could precisely detect the low-concentration antibody. Chi-square test for fourfold table data showed P<0.01. Significant differences in the differences of panel reactive antibody of kidney disease patients were detected between the two methods. Results demonstrated that compared with enzyme-linked immunosorbent assay, Luminex method is more sensitive and accurate, and more suitable for clinical detection.