OBJECTIVE: To improve superparamagnetic iron oxide nanoparticles (SPIO-NP) stability, biocompatibility and tumor-targeting and evaluate their tumor targeting in vitro. METHODS: The folic acid-carboxymethylchitosan-superparamagnetic iron oxide nanoparticles (FA-OCMCS-SPIO-NPs) were synthesized by "three-steps": at first, superparamagnetic oxide iron nanoparticles were synthesized by coprecipitation, then, O-carboxymethyl chitosan and folic acid were successfully immobilized on the surface of SPIO-NPs in turn. The fourier transform infrared spectroscopy, X-Ray diffraction were used to confirm their synthesis, meanwhile, transmission electron microscope (TEM), dynamic light scattering (DLS), Zeta-potential measurement and vibrating sample magnetometry (VSM) were applied to characterize their physicochemical properies. Ferrozine assay and Prussian blue staining were used to evaluate their tumor targeting in vitro. RESULTS: X-Ray diffraction showed the crystalline powder of FA-CMCS-SPIO-NPs agree with the standard Fe3O4, Fourier transform infrared results showed O-carboxymethylchitosan and folic acid were covalently modified on the surface of SPIO-NPs successfully, TEM showed all synthesized SPIO-NPs were almost spherical or ellipsoidal. Their sizes were less then 20 nm, dynamic light scattering (DLS), Zeta-potential results demonstrate the intensity particle size distributions of FA-OCMCS-SPIO-NPs were (41.4±0.132)nm, Zeta potential were (-21.36±15)mV. The surface modification may lead to decrese of magnetisms Ferrozine assay and Prussian blue staining results showed FA-OCMCS-SPIO-NPs had good tumor targeting and the tumor targeting had good relations with the amount of FR on surface of tumor cells. CONCLUSION: FA-OCMCS-SPIO-NPs with strong superparamagnetic property, excellent stability, and good folate receptor targeting is successfully synthesized, which demonstrated the potential for tumor MRI diagnose and therapy.

OBJECTIVE: To synthesize immunomagnetic nanoparticles with uniform particle size, strong superamagnetism as well as strong immune activity which can be specifically and sensitively combined with circulating tumor cells in peripheral blood of patients with breast cancer.METHODS: Superparamagnetic oxide iron nanoparticles containing active carboxyl groups (SMNP-COOH) were synthesized by polyol methods, thermogravimetric analysis was used to determine the amount of carboxyl groups on the surface of SMNP-COOH, while the content of iron was determined by o-phenanthroline. Mediated by 1-ethyl-3,3-dimethylaminopropyl carbodiimide(EDC) and N-hydroxysuccinimide (NHS), immunomagnetic nanoparticles(IMNP) against human breast carcinoma cell line were constructed by binding the monoclonal antibodies against hMAM with SMNP-COOH. X-Ray diffraction was used to confirm their synthesis,meanwhile,transmission electron microscope (TEM), dynamic light scattering (DLS), and vibrating sample magnetometry (VSM) were applied to characterize their physicochemical properties. The conjugation amount of the antibodies and the activity of IMNPs were evaluated by enzyme linked immunosorbent assay (ELISA).RESULTS: X-Ray diffraction showed that the chracteristic peaks of the crystalline powder of SMNP-COOH and IMNP agreed with the Fe3O4 standard. The concentration of iron in SMMP-COOH and IMNP were 0.205 and 0.164 mol•L-1, respectively.TEM showed that both synthesized SMNP-COOH and IMNP were almost spherical or ellipsoidal. The sizes of SMNP-COOH and IMNP were (13.7±3.6) and (15.4±4.5) nm, respectively. Dynamic light scattering(DLS) demonstrated the intensity particle size and polydispersity index (PDI) of SMNP-COOH and IMNP were 23.4 nm and 0.303, and 71.2 nm and 0.175,respectively. VSM results showed that both SMNP-COOH and IMNP had strong superamagnetism, and the saturation magnetization of SMNP-COOH and IMNP were 71.37 and 67.68 emu•g-1Fe, respectively, which confirmed antibody binding may reduce the magnetism of SMNP-COOH. The ELISA results showed the conjugation amount of antibody was about 93 μg on 1 mg SMNP-COOH by covalent bond. The obtained immunomagnetic nanoparticles (IMNP) which were bound with the hMAM monoclonal antibodies could specifically and sensitively combine with breast cancer cell line MDA-MB-415.CONCLUSION: IMNP with strong superparamagnetic property,excellent stability and perfect antibody activity were successfully synthesized, which demonstrate the potential to magnetically separate circulating tumor cells in peripheral blood from patients with breast cancer, thus providing a favorable weapon to accurately detect CTCs in breast tumor patients.

BACKGROUND: Hippocampus is an important region involved in learning and memory. Some methods for freshly isolated hippocampal pyramidal neurons have been described. These methods need multiple enzymes or the procedure is complex.OBJECTIVE: To acquire large quantities of hippocampal neurons from rats suitable for patch clamp study.DESIGN: An animal experimental observation.SETTING: Department of Physiology and Pathophysiology, School of Medicine, Xi'an Jiao Tong University.MATERIALS: The experiments were carried out in the Medical Experimental Center, School of Medicine, Xi'an Jiao Tong University between March and October 2004. The animals were the 10-15-day-old SpragueDawley rats, bothmale and female.METHODS: Hippocampal pyramidal neurons were freshly isolated. Delayed rectifier potassium current and voltage-gated Ca2 + currents were recorded by whole-cell patch clamp configuration.MAIN OUTCOME MEASURES: Acute isolated neurons were observed with a Leica microscope. Recordings of delayed rectifier potassium currents and voltage-gated Ca2+ currents were obtained.RESULTS: ① Under the inverted microscope, the acute isolated neurons had a smooth and glassy surface, a three-dimensional contour, a pyramidal shape with a longer apical dendrite and several basal dendrites. ② After the whole cell recording configuration was formed, voltage-gated barium currents through calcium channels were elicited by 200 ms depolarizing potential from -70 mV to +20 mV, with an increment of 10 mV, a holding potential was -90 mV. Delayed rectifier potassium current (Ikd) was elicited by a protocol consisting of a multiple depolarizing pulses (-60 mV to +50 mV),preceded by a single prepulse step to -50 rmV, with inactivated transient outward potassium currents from a holding potential of -80 mV.CONCLUSION: This method is simple and ideal for the dissociation of neurons from rat hippocampus, and offers a powerful tool for functional analysis of ion channels by patch clamp.

cuses tumors. Survivin seems to be a promising marker for analyzing clinical stages and predicting the prognosis of TCC.

OBJECTIVETo observe the intervention of Shenkangling Decoction (SD) on the renal injury of primary nephrotic syndrome (PNS) children patients of Shen deficiency blood stasis syndrome (SDBSS) and to explore its mechanism.

METHODSTotally 65 PNS children patients were randomly assigned to the combined group (33 cases, treated by SD +Western medicine) and the Western medicine group (32 cases, treated by Western medicine). Meanwhile, 30 healthy children were recruited as the healthy control group from the medical examination center. Those in the Western medicine group were treated with prednisone (5 mg per tablet) at the daily dose of 1.5 -2.0 mg/kg till two weeks after their urine protein turned to negative. Then the dosage was reduced once daily per every other day. The therapeutic course lasted for more than 1 year. For those with no effect of prednisone or partial effect, cyclophosphamide intravenous pulse therapy was additionally applied for 2 successive days per week, a total of 6 times, or they took cyclosporine A. Patients in the combined group additionally took SD while starting treatment of prednisone. SD was decocted in water for oral dose, once daily, taken in two portions until 2 months after prednisone was discontinued. Efficacy was evaluated based on serum levels of chemotactic factor CXCL16, disintegrin metalloproteinase 10 ( ADAM10 ), disintegrin metalloproteinase 17 (ADAM17), albumin (ALB), total cholesterol (TC), and 24-h urine protein excretion (UPE) detected by ELISA before and after treatment.

RESULTSCompared with before treatment in the same group, levels of CXCL16, ADAM10, ADAM17, TC, and 24-h UPE were significantly lower in the two treatment groups (P <0. 01). Compared with the control group, levels of CXCL16, ADAM10, ADAM17, TC, and 24-h UPE significantly increased, and the serum ALB level decreased in the two treatment groups (P <0.01). Compared with the Western medicine group at the same time point, levels of CXCL16, ADAM10, ADAM17, TC, and 24-h UPE significantly decreased in the combined group. The 1 -year recurrence rate and the recurrence times decreased in the combined group (P <0.01). The complete remission rate increased in the combined group (P <0.01).

CONCLUSIONSD could effectively improve the clinical prognosis of PNS children patients possibly by reducing the release of inflammatory mediators such as CXCL16, ADAM10, and ADAM17, decreasing UPE and the TC level, and elevating the serum ALB level.

Objective To study the effect of different dose fractionation on overall survival in patients with limited-stage small cell lung cancer (LS-SCLC). Methods LS-SCLC patients treated with radical combined chemotherapy and radiotherapy (RT) between January 2001 and Dec 2007 were analyzed retrospectively. According to the dose fractionation schemes, patients were divided into three groups:conventional fractionated RT (1. 8 -2.0 Gy,once daily), hyperfractionated RT (1.4 Gy, twice daily) and hypofractionated RT (2. 5 Gy,once daily). Overall survival, disease free survival and pattern of failures of the three groups were compared. A total of 177 patients were enrolled, including 63 patients in conventional fractionated RT group, 79 in hyperfractionated RT group and 35 in hypofractionated RT group. Results The overall follow-up rate was 96. 6%. The patient numbers with follow-up of more than 2 and 5 years were 153 and 92, respectively. The median survival time of the entire group was 22. 4 months, and the 2-and 5-year survival rates were 43.4% and 23. 5%, respectively. The 2-year survival rates for three groups were 31%, 46% and 59% (x2 =7.94,P=0.019), respectively. The 2-year disease free survival for three groups were 20%, 31% and 40% ( x2 = 4. 86, P = 0. 088 ), respectively. In the pairwise comparisons,patients in hypofractionated RT group have better survival than those in conventional fractionated RT group ( x2 = 7. 81, P = 0. 005 ), the effect of hyperfractionated RT group lies between the hypo-and the conventional fractionated RT groups, but no significant differences were detected ( x2 = 2. 31, P = 0. 128; x2 = 2. 95, P =0. 086). The mildest side effect was found in the hypofractionated RT group. No statistically significant differences were found in the patterns of first failure. Conclusion The hypofractionated RT scheme showed potential survival benefits for patients with LS-SCLC and should be considered in the setting of randomized clinical trials.

Objective To compare the efficacies of hepatectomy and liver transplantation for patients with hepatocellular carcinoma (HCC) fulfilling the Milan criteria.Methods From July 2002 to February 2009,121 patients with HCC combined with hepatic cirrhosis fulfilling the Milan criteria were admitted to the Fuzhou General Hospital.Eighty-nine patients who received hepatectomy were in the hepatectomy group,and 32 patients who received liver transplantation were in the liver transplantation group.There were no significant difference in the age,gender,etiology of liver disease,the size of the largest tumor,number of tumors,microscopic venous invasion,microsatellite lesion and tumor differentiation between the 2 groups.The clinical data of the patients in the 2 groups were retrospectively analyzed.The overall survival and disease-free survival were evaluated by Kaplan-Meier method,and differences in survival rates between the 2 groups were determined by Log-rank test.COX proportional hazard was used for univariate and multivariate analysis to evaluate the risk factors for prognosis.Results The median period of follow-up was 37 months.The 1-,3-,5-year survival rates were 86％,63％ and 44％ in the hepatectomy group,and 87％,70％ and 62％ in the liver transplantation group.There was no significant difference in the overall survival rate between the 2 groups (x2 =1.092,P ＞ 0.05).The 1-,3-,5-year disease-free survival rates were 68％,44％ and 26％ in the hepatectomy group,and 80％,65％ and 52％ in the liver transplantation group.There was a significant difference in the disease-free survival rate between the 2 groups (x2 =4.712,P ＜ 0.05).The result of univariate analysis revealed that microscopic venous invasion and microsatellite lesion were significantly correlated with the survival (Wald =9.625,7.340,P ＜ 0.05),and the result of multivariate analysis indicated that microscopic venous invasion was the independent risk factor influencing the survival (Wald =5.008,P ＜ 0.05).Conclusions As for patients with HCC fulfilling the Milan criteria,the overall survival rate of patients who received hepatectomy is not different from those who received liver transplantation,but the disease-free survival rate of patients who received liver transplantation is higher than those who received hepatectomy.Microscopic venous invasion is an independent risk factor influencing the survival.

OBJECTIVE: To explore how to carry out pharmaceutical consultation for anti-infection treatment in neutropenic hematological patients with invasive fugal disease and elaborate the value of clinical pharmacists in anti-infection treatment. METHODS: A total of 41 hematologic malignancies patients with invasive fungal disease who were consulted by clinical pharmacist from October 2014 to June 2016 were enrolled into the study. The etiology, bacteria complication, and infection site were summarized. The other 41 agranulocytosis patients complicated with invasive fungal disease without clinical pharmacist consultation randomly sampled by HIS were used as the control. Statistical analysis were carried out to evaluate the effect of anti-infection treatment. The authors also discussed that as a clinical pharmacist how to carry out pharmaceutical consultation through several typical anti-fungal infection cases. RESULTS: Totally 45 strains of fungi were isolated from the secretion specimens obtained from the 41 patients, including Candida albicans, Candida glabrata, Candidakrusei, Candida tropicalis, Aspergillums, and Cryptococcosis, among which Candida albicans accounted for 60.0%, followed by Aspetgillus (13.3%), Candidakrusei (11.1%), Candida glabrata (6.67%), Candida tropicalisi (6.67%), and Cryptococcosis (2.2%). The main infection site was the lung, followed by the digestive tract and blood stream. The positive rate of bacteria culture was 58.5% among the 41 patients, and the major isolated bacteria were Escherichia colis, Pseudomonas aeruginosa, Enterococcus, and Pseudomonas maltophilia. For the antifungal treatment involving the clinical pharmacists, the cure rate was 48.8%, the significant effective rate was 34.2%, the improved effective rate was 7.4%, the total effective rate of treatment was 72.9%, and the failure rate was 9.3%. There was significant difference in the curative effect between the clinical pharmacist consultation group and the control group (P<0.05). CONCLUSION: The incidence of fungal infection in agranulocytosis patients is high, and most of the patients are complicated with bacteria infection. The most frequently infected site is respiratory tract. Clinical pharmacists can play an important role in the treatment of invasive fungal disease in agranulocytosis patients to ensure the treatment safety and efficacy.

Objective: To investigate the neural connections between Shenmen (HT7)-heart and the brain by observing the tracing viruses co-labeled brain nuclear groups after injection of the pseudorabies viruses (PRV), the reverse transsynaptic virus tracer carrying different fluorescent protein genes, into the myocardium and Shenmen (HT7) point, respectively.Methods: Pseudorabies virus 531 (PRV531) carrying the green fluorescent protein gene and pseudorabies virus 724 (PRV724) carrying the red fluorescent protein gene were injected into the left ventricular wall and Shenmen (HT7) point area of the left forelimb of six C57BL/6 mice, respectively. After 120 h, whole brain tissue was extracted under 4％ paraformaldehyde perfusion to prepare brain sections. Neuronal co-labeling with the tracing viruses was observed under fluorescence microscopy. Results: Co-labeled signals from the mouse ventricular wall and Shenmen (HT7) point region were found at all levels of the mouse central nervous areas, such as the cerebral cortex, hypothalamus, midbrain, pons, and medulla oblongata. The number of co-labeled neurons was higher in the primary motor area, the hypothalamic paraventricular nucleus, the subceruleus nucleus, and the paramedian reticular nucleus. Conclusion: There is a neural connection between Shenmen (HT7), the heart, and the brain, which may be most closely related to the autonomic nervous system.

This study was aimed to establish quality standards of Granati pericarpium and Granati semen. The microscopy and thin layer chromatography (TLC) were used in the identification of Granati semen. According to the 2010 Chinese Pharmacopoeia, the content of moisture, total ash, acid-insoluble ash and alcohol-soluble extract were determined. And the HPLC was used in the content determination of ellagic acid in Granati semen and Granati pericarpium. The results showed that the microscopic characteristics of Granati semen were identified. And TLC identification of Granati semen was established. The content determination method of ellagic acid in Granati pericarpium and Granati semen were established. It was concluded that the established qualitative and quantitative method can be used for quality control of Granati pericarpium and Granati semen.