OBJECTIVE:To facilitate the automatic drug dispensing in hospital pharmacy.METHODS:Full automatic single dose pastill dispensing machine was introduced to hospital central drug dispensing.The running cost was analyzed from two aspects and the feasibility of cutting cost is demonstrated.RESULTS & CONCLUSIONS:The key to achieve automatic drug dispensing in hospital pharmacy is to reduce the running cost to a lowest degree,and the key to lower running cost lie in the omnidirectional controlling of cost,enhancing of performance and reinforcing of the consciousness on cost etc.

Objective To investigate clinical value of three-dimensional transvaginal sonography (3D-TVS) in the diagnosis of septate uterus and the relationship between its parameters and adverse pregnancy.Methods From Mar.2010 to Sept.2011,73 patients (aged 23-35 years) with septate uterus who were diagnosed by 3D-TVS in Fujian Province Maternal and Child Health Hospital were enrolled in this retrospective study.The septum width,septum angle,septum length and the length of remaining uterine cavity were measured among the patients with subseptate uterus,and then,the distortion rate was calculated.The hysteroscopic surgery was used as the gold standard,and the diagnostic titer of 3D-TVS in the diagnosis of septate uterus was determined.Receiver operating characteristic curve (ROC) were plotted to evaluate the diagnostic titer of uterine parameters measured by 3D-TVS in predicting the adverse pregnancy outcome among patients with subseptate uterus.Univariate logistic regression was used to analyze the effectiveness of uterine parameters on adverse pregnancy outcome.Results Using hysteroscopic surgery as the gold standard,the coincidence rate of diagnosis of septate uterus by 3D-TVS was 94％ (69/73).Among the patients with septate uterus,25％ (17/69)were complete septate uterus,75％ (52/69) were subseptate uterus.Among patients with subseptate uterus,the septum length [(2.2 ± 0.6) cm] and distortion rate in patients with adverse pregnancy(0.60 ± 0.10)were significantly higher than those without adverse pregnancy [(1.5 ±0.6) cm,0.43 ±0.13,both P ＜0.05].However,no significant difference in the width,angle and length of septum were observed between the two groups (P ＞ 0.05).The area under ROC curve (AUC) of septum length and distortion rate in determining adverse pregnancy were 0.833 (95％ CI:0.721-0.944) and 0.800 (95％ CI:0.671-0.929),respectively.The optimal cutoff point of septum length was 1.94 cm,with the sensitivity was 74.3％ and the specificity was 76.5％ ; the optimal cutoff point of distortion rate was 0.48,with the sensitivity was 77.1％ and the specificity was 76.5％.The expectation morbidity ratio of adverse pregnancy was 2.717,3.067 and 0.514 by every adding level of septum length,distortion rate,and length of remaining uterine cavity,respectively.Conclusions 3D-TVS showed high accuracy in diagnosis of septate uterus.The septum length and distortion rate may predict the risk of adverse pregnancy,and the value of them can be used for screening adverse pregnancy in clinical practice.

AIM In order to search inhibitors of protein kinase CK2, we observed the inhibitory effects of kaempferol on recombinant human protein kinase CK2 holoenzyme and its kinetics in vitro. METHODSCloning, prokaryotic expression and purification of human protein kinase CK2 α' and β subunits by gene engineering, the two subunits were mixed at equal molar ratio to reconstitute CK2 holoenzyme and identify its biological properties. The CK2 activity was assayed by detecting incorporation of 32P of [γ-32P]ATP into the substrate. The inhibitory effect of kaempferol on CK2 was assayed in the presence of different concentrations of kaempferol. Kinetic analysis of kaempferol-induced inhibition was carried out in the condition that casein concentration was fixed at 2 g·L-1 and ATP was changed at various concentrations(10, 20, 40, 80 μmol·L-1), or ATP was fixed at 10 μmol·L-1 and casein was changed at different concentrations (1, 2, 4, 8 g·L-1). RESULTS Kaempferol was shown to strongly inhibit the holoenzyme activity of recombinant human protein kinase CK2 with IC50 of 1.9 μmol·L-1, which was more effective than chrysin, morin and genistein which are both known as CK2 special inhibitors. Kinetic studies of kaempferol on recombinant human CK2 showed that kaempferol acted as a noncompetitive inhibitor with substrate ATP(Ki=1.1 μmol·L-1) and casein (Ki=3.1 μmol·L-1). CONCLUSIONKaempferol is a novel potent inhibitor of protein kinase CK2 in vitro. Discussions indicate that flavonoid inhibitors of CK2 may adopt different orientations in theactive site of CK2 and that these are determined by the number and position of their hydroxyl groups.

Objective To observe the effect of 7 flavonoids on recombinant human protein kinase CK2 holoenzyme activity and investigate their structure-activity relationship. Methods Recombinant hu-man protein kinase CK2 α' and β subunits were mixed at equal molar ratio to reconstitute CK2 holoen-zyme. The CK2 activity was assayed by detecting incorporation of 32p of [γ-32P] ATP into the substrate for the inhibitory effect by flavonoids and calculation of IC50 was performed according to probability unit (PROBIT) method. Results Myricetin, quercetin, morin, luteolin, kaempferol, apigenin, and chrysin were shown to obviously inhibit recombinant CK2 holoenzyme activity in a concentration-dependent man-ner with IC50 values of 1.18, 0.51, 16.16, 0.86, 1.88, 1.72, and 13.63 umol/L, respectively. Myricetin, quercetin, luteolin, kaempferol, and apigenin were more effective than DRB and A3, which were known as CK2 inhibitors in vitro. Whereas morin and chrysin displayed a similar effect to DRB. Structure-activity study indicated that the major structural requirements for the potent inhibition of CK2 by these flavonoids were hydroxyl group at position 6, 3' and 4'. Different from these requirements, absence of a hydroxyl group at position 3 did not modify their inhibitory potency, while addition of hydroxyl groups at positions 2' or 5' was detrimental to the inhibitory effect on CK2. Conclusion The inhibitory effect of flavonoid on protein kinase CK2 in vitro may be determined by the position of their hydroxyl groups.

Objective To investigete the effect of low-concentration ozone (O3) on voltage-gated calcium channels in fetal rat spinal cord neurons in vitro. Methods The spinal cord neurons were cultured in vitro and identified by immunohistochemistry staining. Then the cells were seeded into the culture dish and randomly divided into 3 groups: control group (group C, n = 5), O3 15 μg/ml (group O3-15, n = 4) and O3 20 μg/ml group (group O3-20, n = 8). The cells were exposed to O3 15 and 20 μg/ml in group O3-15 and O3-20 respectively, while the cells were exposed to air in group C. After 20 min incubation, the electrophysiological activity of calcium channels in neurons was recorded using whole-cell patch-clamp technique. Results Peak calcium current density was significantly increased in group O3-15 and O3-20 compared with group C ( P ＜ 0.05). Half-activation voltage was significantly lower in group O3-15 and O3-20 than in group C ( P ＜ 0.05). Peak calcium current density was significantly higher in group O3 -20 than in group O3-15 ( P ＜ 0.05) . There was no significant difference in half-inactivation voltage among the three groups ( P ＞ 0.05). Conclusion Low concentration of O3 can make the activation of the calcium ion channel easy in fetal rat spinal cord neurons in vitro.

Objective To construct the recombinant plasmid of fusion protein containing respiratory syncytial virus (RSV) cytotoxicity T lymphocyte (CTL) epitope M2:81-95 and heat shock protein (HSP) 70L1, and to investigate its immunogenicity after prokaryotic expression. Methods HSP70L1 gone was cloned from SMMC7721 cells. The M2:81-95 gene fragment was amplified by polymerase chain reaction (PCR) method. Plasmid pET-HSP70L1-M2 : 81-95 (pET-HSP70L1-M2) was constructed, identified and transferred into E. coli BL21 (DE3). Expression of HSP70L1-M2 : 81-95(HSP70L1-M2) was induced by isopropy-β-D-thiogalaetosidc (IPTG). The expressed protein was purified by affinity chromatography and renatured by gradient dialysis. The BALB/c mice were immunized with this fusion protein. IgG antibodies and the subtypes were detected by enzyme-linked immunosorbent assay (ELISA), and CTL responses were determined by methyl thiazolyl tetrazolium (MTT). Results The recombinant plasmid pET-HSP70L1-M2 was successfully constructed. The fusion protein HSP70L1-M2 was expressed in E. coll. The purified protein induced strong RSV-and CTL epitope-specific CTL responses and high titer of protein specific lgG antibody 4.87±0.35. The subtypes were IgG1 (5.53±0.28) and lgG2a (4.40±0.21) and IgG1/ IgG2a ratio was balanced. The titers of lgG, IgG1 and IgG2a in PBS control group were 0.33±0.17, 0.51±0.21 and 0, respectively, which werc significantly lower than those in immunized group (t = 3.512, 3.681, 5.856; P<0.05). Conclusions The recombinant plasmid pET-HSP70L1-M2 is successfully constructed and the fusion protein is expressed and purified. HSP70L1-M2 induced strong RSV-and CTL epitope-specific CTL responses and mixed T helper cell (Th)1/Th2 response in BALB/c mice.

Object To study the chemical constituents of a new species belonging to genus Tinospora (Menispermaceae)— the Tinospora hainanensis H S Lo et Z X Li Methods Isolation and purification were carried out on silica gel column, identified by physico chemical properties and structurally elucidated by spectral analysis Results 4 non alkaloids were obtained They were 24 epi makisterone A (Ⅰ), octacosanoic acid (Ⅱ), octacosyl alcohol (Ⅲ) and hexacosyl alcohol (Ⅳ) Conclusion All of the 4 compounds were obtained from this plant for the first time, and compounds Ⅰ, Ⅱ and Ⅳ were obtained from genus Tinospora for the first time

Objective To investigate the relationship between the efficacy of autologous cultured melanocyte transplantation and serum levels of interleukin-17 (IL-17) and FoxP3 in patients with vitiligo.Methods Forty patients with stable vitiligo vulgaris were included in this study,and received autologous cultured melanocyte transplantation.Six months after the transplantation,treatment efficacy was evaluated,and patients were classified into the successfully treated group (n =25) and unsuccessfully treated group (n =15).Peripheral blood was collected from all the patients before the transplantation,and enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum levels of IL-17 and FoxP3.Statistical analysis was done using two independent samples t-test with the SPSS software (version 17.0).Results The successfully treated patients showed lower serum levels of IL-17 ((15.29 ± 7.86) vs.(43.88 ± 13.02) ng/L,P ＜ 0.05),but higher serum levels of FoxP3 ((6.08 ± 2.03) vs.(3.37 ± 1.81) ng/L,P ＜ 0.05) than the unsuccessfully treated patients.Conclusion The increased serum IL-17 and decreased serum FoxP3 may contribute to the failure of autologous cultured melanocyte transplantation in patients with vitiligo.

Objective To observe the ultrastructural features of recessive dystrophic epidermolysis bullosa inversa(RDEB-Ⅰ)and to detect the mutations of COL7A1 gene in a family with RDEB-Ⅰ.Methods A 24-year-old male patient complained of recurrent vesicles in the skin for 24 years.The lesions began as generalized pruritic vesicles and bullae soon after birth,with a predilection for areas subject to friction,and showed a trend to be worse in summer but mild in winter.No photosensitivity was observed.When he was 3 to 4 years old,the lesions were decreased in number,with the only involvement of the trunk and abdomen;thereafter,the lesions were improved year by year.The patient suffered from nephritis at the age of 5 years,which progressed into renal failure at the age of 15 years.He received renal transplantation and was given long-term oral tacrolimus and mycophenolate mofetil,which leaded to an improvement in the lesions.The family history was unremarkable,and the marriage between her parents was not consanguineous.Dermatological examination revealed large areas of irregularly-marginated,hypopigmented,atrophic scar on the waist,back and abdomen with onychodystrophy involving multiple nails.No vesicles were observed.Immunofluorescence antigen mapping and transmission electron microscopy were conducted to observe the expression of type Ⅶ collagen in and ultrastructure of cutaneous lesions from the patient.Venous blood samples were obtained from the patient as well as his parents and 3 sisters,and drill biopsy specimens were obtained from the margin of vesicular lesions and unaffected anterior tibial skin of the patient.DNA specimens were obtained from the blood samples of the family members and 150 unrelated healthy controls,and RNA was extracted from the biopsy samples of the patient.PCR and direct sequencing were carried out to detect mutations in COL7A1 gene,and reverse transcription-PCR was conducted to confirm the mutation at mRNA level.Results Skin cleavage was observed under lamina densa in the dermis,with a decrease in anchoring fibrils and expression of type Ⅶ collagen in the lesions of the patient.A heterozygous synonymous mutation c.C5499T which created a new splicing site and leaded to a premature terminal codon,as well as a heterozygous missense mutation c.C6205T(C-T transition at codon 2069:CGT to TGT)which leaded to the substitution of arginine by cysteine,were identified in the COL7A1 gene of the proband and all of his sisters,but not in any of the unrelated controls.The c.C5499T and c.C6205T mutations were inherited from the patient's father and mother respectively.Conclusion The compound heterozygous mutations c.C6205T and c.C5499T may be responsible for RDEB-Ⅰ in this patient.