AIM: To determine 5 different aphrodisiacs,including apomorphine,yohimbine,vardenafil,sildenafil and alprostadil,adulterated into Chinese proprietary medicine by LC/MS(n),and build a laboratory library including MS(n) spectra and retention time. METHODS: ZORBAX SB C_18 column(150 mm?4.6 mm,5 ?m)was applied.The mobile phase consisted of acetonitrile and 0.5% formic acid changing by linear gradient elution,and the flow rate was 1.0 mL/min.The ion trap mass spectrometer with positive ion mode electrospray(ESI?) was applied. RESULTS: In the mass spectrum apomorphine,yohimbine,vardenafil and sildenafil showed abundant pseudomolecular ion +,and alprostadil showed abundant pseudomolecular ion -.All the MS(n) spectra showed characteristic product ion. CONCLUSION: It is proved that the method is simple,reliable and suitable to determin aphrodisiac adulterated to health food.

AIM:To establish a specific method for the identification of 26 chemical drugs added illegally into analgesic-antipyretic traditional Chinese medicines and health food. METHODS: The liquid chromatography-ion trap mass spectrometry method was used.Comparing with retention time and spectrums of references in library set up by ourselves,the target compounds in sample were screened and identified. RESULTS: Sixty samples were tested and Naproxen,Ibuprofen and Aspirin were detected out. CONCLUSION: The method is accurate,sensitive and easy to operate, which is first reported in China and so many compounds could be detected at the same time.

Quality control of traditional Chinese medicine (TCM) has become bottlenecks of TCM industry devel-opment and TCM international recognition. To solve the retational problems, we explored the establishment of new quality control method based on efficacy and safety. Firstly, material basis of TCM efficacy was investigated deeply and systematically. Then, quality control method based on efficacy was established by using active ingre-dients as markers. We also establish detection methods based on safety, such as pesticide residues, heavy metals and harmful elements , mycotoxins .

Objective To analyze the medical service capacity of primary healthcare in Zhejiang province since the ongoing healthcare reform and put forward suggestions. Methods Key indicators of healthcare resources and medical service utilization from 2009 to 2015 reflecting the primary healthcare were identified,for a quantitative analysis in terms of the structure-process-outcome dimensions. Results In terms of structural service capacity,the average headcount growth rate of primary healthcare′s technical personnel was 5. 7% per year; the personnel competence structure kept improving; the hospital beds at primary institutions and their ratio among all were slightly decreased,with better devices and informatization. In terms of procedural service capacity,the proportion of primary institutions with contract-based services amounted to 89. 9%,with the standard contract signing rate up to 18. 8%. In terms of consequential service capacity,the average growth rate of the number of outpatient visits at primary institutions was 6. 3%. The amount and proportion of inpatients were slightly decreased,while the hospital bed utilization ratio was increased slightly. Conclusions It is necessary to further strengthen the training and introduction of primary healthcare professionals. The functional orientation of hospitals at various levels should be clarified, encouraging contract-signing of general practitioners, promote the medical insurance payment reform featuring the capitation payment at primary level,and improve the income distribution and incentive mechanism.

The drugs such as sildenafil adulterated in herbal products and dietary supplements may endanger human health. The number of the new modified derivatives is increasing recently. Based on Q-TOF-MS, a new sildenafil analogue was found. It was isolated and purified by preparative liquid chromatography. Its structure was determined by NMR, as 1-[4-propoxy-3-(6, 7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo[4, 3-d] pyrimidin-5-yl)phenylsulfonyl]-4-methylpiperazine. Compared with sildenafil, the ethoxy group of the benzene ring moiety was moved to the propoxy group, which had not been reported in China. The mass spectrometric behavior pattern of the structure type was summarized, which can greatly accelerate the structural analysis of novel analogues.
Chromatography, High Pressure Liquid ; Dietary Supplements ; analysis ; Drug Contamination ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Piperazines ; chemistry ; isolation & purification ; Sildenafil Citrate ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To evaluate the in vitro activity of seven imidazole antifungals against clinical isolates of common Candida species.Methods According to the Clinical and Laboratory Standards Institute (CLSI) microdilution method M27-A3,the in vitro activity of luliconazole,ketoconazole,miconazole,econazole,clotrimazole,sertaconazole and bifonazole was determined among 183 clinical isolates belonging to 5 species of Candida.Results The minimal inhibitory concentration range was 0.03-8 (geometric mean:0.067) mg/L for ketoconazole,0.03-16 (geometric mean:0.071 ) mg/L for miconazole,0.03-8 (geometric mean:0.207) mg/L for econazole,0.03-8 (geometric mean:0.061 ) mg/L for clotrimazole,0.03-16 (geometric mean:0.187) mg/L for sertaconazole and 0.03 -＞16 (geometric mean:1.050) mg/L for bifonazole. Luliconazole exhibited a superior activity against the 5 species of Candida in vitro,with the MIC range being 0.03-8 mg/L,geometric mean MIC 0.087 mg/L,MIC50 0.06 mg/L and MIC90 0.5 mg/L,respectively.However,some Candida isolates were identified to be relatively insensitive to these tested antifungals,including luliconazole.Conclusion All the tested imidazole antifungals,except for bifonazole,show an excellent activity against Candida species in vitro,but there exist a few Candida strains with relative insensitivity.

Objective To investigate the effects of Candida albicans on the expression of tumor necrosis factor-α(TNF-α)and activation of the intracellular signaling molecule p38 mitogen-activated protein kinase(p38MAPK)in a human acute monocytic leukemia cell line THP-1. Methods Some THP-1 cells were divided into several groups in vitro: two C. albicans groups treated with 105 CFU/ml and 106 CFU/ml heat-killed C. albicans respectively, a lipopolysaccharide (LPS)group treated with 100 μg/L LPS, a blank control group treated with RPMI 1640 medium, two dexamethasone-inhibited groups pretreated with 40 μg/L dexamethasone for 30 minutes followed by treatment with 106 CFU/ml heat-killed C. albicans and LPS respectively. After treatment for 1, 3 and 6 hours, real-time fluorescence-based quantitative PCR was performed to measure TNF-α mRNA expression in THP-1 cells in the above groups. Enzyme-linked immunosorbent assay(ELISA)was conducted to determine the level of TNF-α protein in the supernatant of THP-1 cells treated with 106 CFU/ml heat-killed C. albicans, 100 μg/L LPS or RPMI 1640 medium(blank control group)for 24 hours. Western blot was performed to measure the protein expression of p38MAPK and phosphorylated p38MAPK in THP-1 cells after treatment with 106 CFU/ml heat-killed C. albicans or RPMI 1640 medium (blank control group)for 30 and 60 minutes. Statistical analysis was carried out by using two-way analysis of variance, one-way analysis of variance and the least significant difference(LSD)-t test. Results Significant differences were observed in the mRNA expression level of TNF-α among the C. albicans groups, LPS group and blank control group (F = 110.98, P < 0.001). The mRNA expression level of TNF-α in THP-1 cells increased over time in a time-dependent manner after C. albicans treatment, with significant differences among different time points (F = 701.680, P < 0.001). Compared with the blank control group, both 106-CFU/ml C. albicans group and LPS group showed a significant increase in TNF-α protein expression (6385.70 ± 533.99 ng/L and 3212.06 ± 353.00 ng/L vs. 147.10 ± 0.53 ng/L, P < 0.001 and 0.005, respectively). An obvious increase was observed in the expression level of phosphorylated p38MAPK protein, but no significant changes were noted in that of p38MAPK protein, in THP-1 cells treated with 106 CFU/ml C. albicans for 30 and 60 minutes compared with the blank control group. The mRNA expression level of TNF-α significantly decreased in dexamethasone-pretreated 106-CFU/ml C. albicans group and LPS group compared with those without dexamethasone pretreatment(3.77 ± 0.62 vs. 208.50 ± 10.50, 6.20 ± 1.93 vs. 161.35 ± 1.65, both P < 0.001). Conclusions Heat-killed C. albicans can induce the activation of p38MAPK in and secretion of TNF-α by human THP-1 cells, which then participate in the innate immune response against C. albicans.

The control handles( including financing, provider payment, organization, regulation and behavior) proposed by World Bank in health policy research as the basic framework,and the nature and positioning of family doctors′ contract-based service as the precondition, we recommended the system framework of such a mechanism, comprising one contract, three regulations, five control handles and seven supporting policies. Such a framework can serve as the cornerstone for family doctors′ contract-based service experiment in Zhejiang province for the sake of hierarchical medical system.

Objective To evaluate the expression of miR-196b in newly diagnosed pediatric acute myeloid leukemia (AML) and its clinical signiifcance. Methods Fifty-two AML children were enrolled in this study and 30 non-leukemia com-pared children were selected as controls. The expressions of miR-196b were detected in bone marrow samples by real-time quan-titative PCR (q-RT-PCR) and the results were expressed in 2-??Ct. Results miR-196b expressions were signiifcantly higher in M4-5 and lower in non-M4-5 of AML children than those in control (P<0.01), with a lowest level in t (15;17) and a highest level in MLL subtypes (P<0.01). The miR-196b expressions were signiifcantly different among different prognosis groups (P<0.01) and the level in the favorable prognostic group was lower than in poor prognosis group. It was also found that miR-196b expres-sion was lower in remission group than that in no-remission group after the ifrst induction remission therapy (P<0.05). Mean-while, the expression of miR-196b in the children with WBC≥100×109/L were statistically higher than that in the children with WBC<100×109/L (P<0.01), and miR-196b level was positively correlated with the platelet counts (r=0.302, P=0.030). Conclu-sions miR-196b expression is increased in poor prognosis group of AML children, and high expression of miR-196b is related with low response rate and poor prognosis. miR-1966 is expected to become a new target for the treatment of AML.

Objective To investigate whether human neutrophils kill Candida albicans through recognition of insoluble β-glucan in cell walls of C.albicans (CalG) by dectin-1,a C-type lectin receptor.Methods Neutrophils were obtained from peripheral blood of healthy human subjects and cultured in vitro.Real-time PCR was carried out to quantify the mRNA expressions of dectin-1 and Toll-like receptor 2 (TLR2) in neutrophils challenged with CaIG of 100 mg/L for 1,6,and 24 hours.A Fluoro hydrogen peroxide (H2O2) detection kit was used to determine H2O2 levels in some neutrophils exposed to CaIG (100 mg/L) for 15 minutes,2 hours,6 hours,as well as in some neutrophils preincubated with laminarin (a dectin-1 inhibitor) of 100 mg/L and 50 mg/L for 30 minutes followed by challenge with CaIG of 100 mg/L for 2 hours.Colony forming units (CFUs) were counted after the incubation of C.albicans with neutrophils pretreated with laminarin of 100 mg/L and 50 mg/L for 30 minutes.Results The relative mRNA expression level of dectin-1 was 2.8195 + 0.1669,5.4859 + 0.7244 and 3.6041 + 0.5372 in neutrophils challenged with CaIG for 1,6 and 24 hours,respectively,significantly higher than that in unchallenged neutrophils at these corresponding time points (all P ＜ 0.01).The level of H2O2 was (64.55 + 15.67),(290.34 + 30.56),and (208.54 ± 26.88) μ mol/L respectively in neutrophils treated with CaIG for 15 minutes,2 hours,and 6 hours respectively,compared to (22.05 ± 3.99) μmol/L in untreated neutrophils (all P ＜ 0.01).The pretreatment with laminarin of 100 and 50 mg/L attenuated the release of H2O2 in CaIG-treated neutrophils by 73％ ((80.45 + 22.41) μ mol/L,P＜ 0.01) and 45％ ((130.42 + 44.55) μmol/L,P＜ 0.01),respectively,compared with neutrophils treated with CaIG only.The fungicidal activity of neutrophils against C.albicans was also significantly inhibited by pretreatment with laminarin of 50 and 100 mg/L (both P＜ 0.01).Conclusions Dectin-1 may be involved in the secretion of H2O2 as well as killing of C.albicans by human neutrophils,which may provide a preliminary evidence for adoptive transfer of neutrophils as an approach to the treatment of systemic C.albicans infection.