Objective To establish a method for the chiral separation and determination of S-isomer in epinephrine hydrochloride injection by HPLC with chiral mobile phase additives. Methods Column of Purospher? STAR RP-18 (4. 6 mm×250 mm, 5 μm) was used. The mobile phase was acetonitrile-10 mmol·L-1 sodium dihydrogen phosphate buffer containing 10 mmol·L-1 sulfobutylether-b-cyclodextrin (pH adjusted to 3. 0 with phosphoric acid) (98. 5:1. 5), detection wavelength was 280 nm, the flow rate was 0. 8 mL·min-1 , and the column temperature was 30 ℃ . Results Good linear relationship was established between the peak area and the concentration of S-isomer over the range of 5. 02-1501. 50 μg·mL-1 (R2 =0. 999 7). The detection limit was 0. 05 μg·mL-1 . Conclusion The proposed method shows high repeatability and durability. It can be employed for the quality control of S-isomer in epinephrine hydrochloride injection.

Objective To identify the pathogenic gene for a Chinese Han consanguineous marriage family with autosomal recessive cerebellar ataxia by homozygosity mapping and mutation analysis.Methods Six members of the family were enrolled in this study,including 3 patients,the unaffected sibling and their parents of first cousin marriage.After excluding GAA repeats mutation of FXN gene,whole-genome single nucleotide polymorphism (SNP) microarray scanning and homozygosity mapping were performed to localize the candidate gene.The coding regions and intronic flanking sequences of the candidate genes were analyzed.Results Four candidate regions were identified,including 2p25.3,9q22.2-34.3,13q12.3-14.3 and 17p13.The SETX gene localizing in 9q22.2-34.3 that is responsible for ataxia with oculomotor apraxia 2 was analyzed at first.There were 4 mutations in exon 10,including three missense mutations (c.3576T ＞ G,p.D1192E ; c.3754G ＞ A,p.G1252R; c.4156A ＞ G,p.I1386V) and a deletion mutation (c.5084_5087delAGTC,p.Q1695_S1696del).Three patients were homozygous of the 4 mutations,an unaffected sibling was normal,and their parents were heterozygous of 4 mutations.Conclusions The pathogenic haplotype comprising four mutations of the SETX gene was identified in the consanguinity family.c.5084_5087delAGTC (p.Q1695_S1696del) is a novel mutation.The affected individuals of this family were characterized by mild phenotype and slow progress without oculomotor apraxia,indicating the clinical variability of the disease.

Objective To investigate the clinical features and genetic mutations of spinocerebellar ataxia type 17 (SCA17).Methods The pathological CAG triplet repeat expansions of the SCA3,SCA1,SCA2,SCA6,SCA7,SCA8,SCA12,SCA17 and dentatorubral pallidoluysian atrophy genes were analyzed in 708 probands of autosomal dominant familial SCA and 1 19 sporadic SCA cases.The CAG repeats of TATA-binding protein (TBP) gene were amplified by means of polymerase chain reaction and agarose gel electrophoresis.For the samples with two alleles,fragment analysis based on CEQ8000 sequencer was applied to analyze the CAG repeat numbers.Furthermore,the correlation between clinical features and CAG repeat in the TBP gene was studied carefully.Results The expanded CAG repeats in the TBP gene was detected in 5 cases with 37/50,36/45,38/52,38/53,36/54 separately.And the main clinical manifestations were ataxia and memory impairment.Conclusion These findings indicate that SCA17 might be a rare subtype of SCA in the Chinese population and the clinical features of SCA17 cover a wider spectrum than previously reviewed.

Objective:To establish a method for the simultaneous determination of chlorphenamine maleate and ephedrine hydro-chloride in chlorphenamine maleate and ephedrine nasal drops by HPLC. Methods:An HPLC method was performed on a column of Purospher STAR RP18(150 mm × 4. 6 mm,5 μm)with the mobile phase of acetonitrile -potassium dihydrogen phosphate buffer(pH 3. 0 ± 0. 1)(18:82 v/v)at the detection wavelength of 210 nm(adjusting the flow rate and column temperature to 1. 0 ml·min-1 and 35℃,respectively). The injection volume was 20 μl. Results:A good linear relationship was established between the peak response and the concentration of ephedrine hydrochloride and chlorphenamine maleate over the range of 19. 81-118. 85μg·ml-1(r=0. 999 6) and 6. 21-37. 25 μg·ml-1(r=0. 999 8),respectively. The mean recovery of ephedrine hydrochloride and chlorphenamine maleate was 100. 39%(RSD=0. 69%,n=9)and 100. 11%(RSD=0. 60%,n=9),respectively. Conclusion:The proposed method shows high repeatability,good durability and promising accuracy. It can be employed for the determination of two components in chlorphena-mine maleate and ephedrine nasal drops.

Objective To explore the peripheral blood microRNA (miRNA) expression of patients with gastric cancer,and to establish specific peripheral blood miRNA expression profile of gastric cancer,which would provide the evidencc for investigating the role of miRNA in the genesis and development of gastric cancer and looking for new molecular markers of gastric cancer.MethodsA total of 6 gastric cancer patients and 6 healthy volunteers were selected.The totat RNA of peripheral blood was extracted for miRNA expression profile examination and hioinformation analysis. The results of microarray were verified by real-time PCR. The online miRNA target gene pr(e)diction software was used to predict and screen miRNA differentially expressed target genes. Results Compared with control group,there were 54 differentially expressed miRNA in gastric cancer group,of which the expression of 35 miRNA (miRNA-504,mi RNA-183,miRNA- 938,miRNA-1285,miRNA- 576-3p,miRNA-663,etc) were up-regulated and 19 miRNA (miRNA-433,miRNA-193b,miRNA-329,miRNA 409-3p,miRNA 154,el(e)) were down-regulated.The results of real-time PCR indicated that there was a good consistency between PCR verificd results and microarray results in 2 up-regulated miRNA (miRNA 504 and miRNA-183) and 2 down-regulated miRNA (miRNA-443 and miRNA- 193b).ConclusionThere is specific peripheral blood miRNA expression profile of patients with gastric cancer,and these differentially cxpressed miRNA will likely become new diagnostic biomarkers of gastric cancer.

BACKGROUND: Orphanin, which was found in recent years, is an important bioactive polypeptide. Its extensive distribution in the central nervous system and peripheral tissue suggests a function of adjusting global behaviors.OBJECTIVE: To explore the correlation of plasm orphanin level and monoamine neurotransmitters in patients with postpartum depression by determining the levels of orphanin and monoamine neurotransmitters such as 5-hydroxytryptamine (5-HT) and dopamine(DA) in patients.DESIGN: Case-control experiment taking diagnosis as evidence.SETTING: Departments of Obstetrics and Gynecology, Changzheng Hospital and Changhai Hospital, Second Military Medical University of Chinese PLA.PARTICIPANTS: Twenty-five healthy women and seventeen women patients with postpartum depression after delivery who came in the Department of Obstetrics and Gynecology, Changzheng Hospital and Changhai Hospital, Second Military Medical University of Chinese PLA for further consultation during February 2002 to October 2004 were enrolled in this study, and served as control group and postpartum depression group, respectively. All the enrolled women, aged 21 to 43 years, with fair education levels, and voluntarily participated in this study. Women patients of 15 to 25 days after delivery suffered from the incipient postpartum depression with total scores of Edingburgh Postnatal Depression Scale (EPDS) above 13. Informed consents were obtained from all the subjects.METHODS: The levels of orphanin and monoamine neurotransmitters in venous blood from 25 healthy women and 17 women patients with postpartum depression were determined. ① Sample collecting: Blood was taken from women in the postpartum depression group 2 weeks after onset, while from control group 35 days after delivery. ② Assay method:The level of orphanin was determined with radioimmunoassay (RIA). The reagent kit was provided by phoenix company (USA) and the procedure was strictly performed according to the instructions on the reagent kit. The levels of 5-HT and DA were measured with high-performance liquid chromatography or electrochemical detection method.MAIN OUTCOME MEASURES: ① Changes in the levels of orphanin as well as 5-HT and DA in postpartum depression group and control group. ② Correlation of the total scores of EPDS and the levels of orphanin in postpartum depression group.RESULTS: Twenty-five healthy women and seventeen women patients with postpartum depression after delivery all participated in the result analysis. ① The level of orphanin in the postpartum depression group[(27.39±6.04) ng/L] was significantly higher than that in the control group [(10.37±3.65) ng/L](P ＜ 0.01),the levels of 5-HT in the postpartum depression group [(0.93±0.21) μmol/L] was remarkably lower than that in the control group[(1.43±0.36) μmol/L](P ＜0.05) ,and the levels of DA in postpartum depression group[(2.15±0.41) μmol/L] was obviously lower than that in the control group [(3.64±0.72) μmol/L]( P ＜ 0.01). ②The level of orphanin was negatively correlated with those of 5-HT and DA in postpartum depression group(r = -0.601 , -0.593,P ＜ 0.05). ③ The total scores of EPDS were significantly positively correlated with the level of orphanin (r = 0.512, P ＜ 0.05), and negatively correlated with the levels of 5-HT and DA (r = -0.571,-0.526, P＜ 0.05).CONCLUSION: The level of orphanin in patients with postpartum depression is increased and is negatively correlated with the levels of 5-HT and DA, but positively correlated with the total scores of EPDS.

Objective To analyze the histological changes of bone diseases and to investigate the noninvasive measurements for diagnosing renal osteodystrophy (ROD) in maintenance dialysis patients . Methods Ninety-one patients were selected to receive bone biopsy . The bone samples were stained with HE, toluidine blue and Masson, and were examined with light microscopy . The levels of immunoreactive parathyroid hormone (iPTH), osteoprotegerin (OPG),sRANKL and osteocalcin (OCN) were determined in the patients enrolled from 2004 to 2006 . The level of iPTH was measured by radioimmunoassay . OPG and sRANKL were measured by ELISA,and OCN was measured by chemiluminescence . Results The incidence of ROD in the maintenance patients was 100% . According to the histological appearance, 50 cases (54 .9%) were high turnover bone disease (secondary hyperparathyroid bone disease), 9 cases (9 .9%) were low turnover bone diseases(osteomalacia and adynamic bone disease), and 32 cases(35 .2% ) were mixed bone disease . The level of iPTH in patients with ROD was significantly increased compared with healthy controls . It was the lowest in low turnover bone diseases . There was no difference among three types of ROD . OPG level was significantly increased compared with healthy controls [(2176 .58±1576 .08) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . The level in high turnover bone diseases was higher than that of the healthy controls [(2261 .85±1712 .22) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . There was no difference among three types of ROD .sRANKL level in high turnover bone disease was significantly increased compared with healthy controls [(0 .328±0 .524)pmol/L vs (0 .084±0 .190) pmol/L, P＜0 .05] . OCN level was also higher than that of the healthy controls (P＜0 .05), and the OCN level in low turnover ROD was the lowest among three types of ROD . OCN level in mixed ROD was dramatically increased as compared to low turnover ROD [(226 .633±66 .455) pmol/L vs (193 .03±104 .269) pmol/L, P ＜0 .05] .Conclusions The histological changes of bone disease can be indicated by iPTH level, but the types of ROD can not be distinguished according to iPTH level neither be differentiated by the levels of OPG, sRANKL and OCN . Bone histomorphometry is still the golden standard for diagnosing renal osteodystrophy .

Objective To investigate the management and control of hypertension in patients with chronic kidney disease(CKD) and its associated factors.Methods Data of 726in-patients with CKD and hypertension who hospitalized in our hospital from March 2009 to April 2010 were studied.Results 91.74％ of patients was treated with antihypertensive medications,and 21.21％, 22.59％, 19.56％, 28.37％ of patients received 1, 2, 3, ≥4 antihypertensive drugs,respectively.42.4％ of patients with CKD and hypertension could be controlled up to the standard,and the mean blood pressure was(137.86±20.75)/(76.30±11.35) mm Hg.There was significant difference among stage 1 plus 2, 3, 4 plus 5 (non-dialysis), 5 (dialysis) kidney diseases, with the hypertension control rate being 50.8％, 46.7％, 42.0％, 33.5％, respectively.The hypertension control rate of non-dialysis patients was significantly higher than that of dialysis (44.9％ vs 33.5％,P＜0.05).There was no significant difference between blood dialysis group and peritoneal dialysis group(32.3％ vs 38.7％, P＞0.05).Multivariate Logistic regression analysis showed that female (OR=1.787, 95％CI 1.045-3.056)and ACEI application (OR=4.378, 95％CI1.830-10.472) were positively associated with hypertension control.Whereas, diabetes (OR=0.415, 95％CI 0.188-0.919)and pulse pressure (OR =0.847, 95％ CI 0.811-0.885) were associated with inadequate blood pressure control.ConclusionsDespite almost universal hypertension treatment is used in patients with CKD and high blood pressure, the hypertension control rate is still suboptimal.Female and ACEI are positively associated with adequate hypertension control, whereas diabetes and pulse pressure are negatively associated with the standard.

Objective To estimate the application value of a standard operating procedure (SOP) in the detection of syphilitic anticardiolipin reagin. Methods Clinical laboratories from 9 local hospitals in Shanghai participated the program. Quality control samples with unknown target value were qualitatively and quantitatively examined according to the uniform SOP in these laboratories with the same reagent and facility of horizontal reaction. External quality assessment (EQA) was carried out by using seven serum samples with no, or low (1∶ 128 dilution) to high (1∶1 dilution) concentrations of target before and after the implementation of SOP. The test results were statistically analyzed and the reasons for the detecting error were assessed. Results A total of 388 tests were performed in the 9 clinical laboratories. The total accuracy rate was 93.0%, including 40.2% in the detection of samples with 1 ∶ 8 dilution of target, 49.2% in the detection of samples with 1 ∶ 16 dilution of target, and 3.6% in the detection of samples with 1 ∶ 32 dilution of target. No forward bias was observed in these tests. There was a significant difference in the accuracy rate between the two times of EQA before and after the implementation of SOP (x2 = 4.17, P < 0.05). Conclusions The improved standard procedure for nontreponemal antigen test is beneficial to the decrease of testing error, and may provide a basis for the establishment of SOP and implementation of internal quality assessment.