Objective:To evaluate curative impact of booster immunization with paternal lymphocytes on recurrent spontaneous abortors(RSA)with less reaction of primary patermal lymphocyte immunization.Methods:RSA patients with insufficient materno-fetal immuno-recognition were selected by flow cytometry of blocking antibody analysis and immunized with either induced paternal lymphocytes pretreated by IFN-? in vitro to those of anti-CD3-BE and anti-CD4-BE Ab lower than 0% or direct intradermal vaccination with their paternal lymphocytes without IFN-? pretreatment to those of anti-CD3-BE Ab beyond 0%.Reassessment of blocking antibodies was performed at the end of the second immnunization course.Results:Levels of blocking antibodies were significantly raised after the secondary booster immunization in RSA with insufficient materno-fetal immnuno-recognition whose blocking antibodies continuously decreased after being treated by the primary paternal lymphocyte immunization.No improvement of parameters was observed except the blocking effect in patients receiving secondary direct intradermal vaccination treatment.Conclusion:It is necessary for the RSA with insufficient materno-fetal immuno-recognition to experience secondary booster immunization preferably with paternal lymphocytes pretreated by IFN-? in vitro.

Objective To establish a sensitive, rapid and simple high-performance liquid chromatography (HPLC) method for the determination of amphotericin B (AraB) and itraconazole (ITZ) in human cerebrospinal fluid (CSF). Methods Solid-phase extraction (SPE) was involved in sample preparation in this method, followed by reverse-phase separation under two different chromatographic systems. Results The tinearity for both AraB and ITZ ranged from 5 ng/mL to 100 ng/mL and the inter-day and intra-day RSD were both below 7.6% at the concentrations of 20,50 and 100 ng/mL. Conclusions The method we established has been applied to the therapeutic drug monitoring on cryptococcal-meningitis-infected patients who were given comedication of these two drugs.

Objective:To investigate the protective effect of schisandrin B (SchB)on the cerebral ischemia reperfusion injury of the rats and the influence in HSPA12B/PI3K/Akt signaling pathway,and to explore the mechanisms.Methods:130 SD rats were divided into sham group,cerebral ischemia reperfusion injury model group (model group),low dose of SchB group (SchB 3 mg· kg-1 ,SchB1 group),middle dose of SchB group (SchB 10 mg·kg-1 ,SchB2 group)and high dose of SchB group (SchB 30 mg·kg-1 ,SchB3 group)(n=26).The rats in sham group didn’t plug lines;the rats in model were used to establish ischemia reperfusion models;the rats in SchB1 ,SchB2 and SchB3 groups were firstly pretreated with different doses of SchB for 7 d,and then they were used to build cerebral ischemia reperfusion injury models.The nerve dysfunction of rats was evaluated by neurologic deficit score.The cerebral edema was detected by measuring the content of water in brain tissue.The morphological changes of brain tissue were observed by toluidine blue staining.The levels of nuclear factor kappa B (NF-κB), tumor necrosis factor-α(TNF-α),interleukin-1 (IL-1)and interleukin-6 (IL-6)in the brain tissue were detected by ELISA.Western blotting method was used to detect the protein expression levels of heat shock protein A12B (HSPA12B ), serine-threonine kinase (Akt ) and phosphorylation serine-threonine kinase (p-AKT ). Results:Compared with sham group,the neurologic deficit score of rats in model group was significantly increased (P <0.01),and the content of water in brain tissue was increased (P < 0.01 );the brain tissue structure was loosened,and the mesenchyme appeared edema;the NF-κB,TNF-α,IL-1,and IL-6 levels were increased (P <0.01),and the expression levels of HSPA12B and p-Akt proteins were decreased (P <0.01).Compared with model group,the neurologic deficit scores of the rats in SchB1 ,SchB2 ,and SchB3 groups were decreased (P <0.01),and the contents of water in brain tissue of the rats in SchB2 and SchB3 groups were decreased (P <0.05);the edema of nerve cells was alleviated,and the cavities were reduced;the NF-κB,TNF-α,IL-1,and IL-6 levels were decreased (P <0.05 or P <0.01),the expression levels of the HSPA12B protein in SchB2 ,and SchB3 groups were increased (P <0.05),and the p-Akt protein expression levels of the rats in SchB1 ,SchB2 ,and SchB3 groups were increased (P <0.01).Conclusion:SchB could protect the cerebral ischemia reperfusion injury of rats,its mechanism may be related to regulating the HSPA12B/PI3K/Akt signaling pathway and inhibiting the inflammatory reaction damage to the nerve cells of reperfusion.

Clinical internship is for bachelor students to learn how to integrate theories in textbooks with clinical practice.Clinical pharmacist training is for pharmacists to obtain and enforce the capability of rational use of drugs.These two are at the different stages of clinical pharmacist cultivation mode.They are different in teaching nature,goal and students' background.The necessity of clinical internship bridged to clinical pharmacist training is discussed in this paper and after the convergence,the teaching design of chnical internship program bridged to the training is characterized by basic education which focuses on basic theories,knowledge and skills of clinical medicine and clinical pharmacy relevant to a certain common disease.The optimized internship program will work as a good foundation for students to face the challenges of clinical pharmacist training after graduation from school.

ObjectiveTo investigate the expression of heme oxygenase-1 (HO-1) in lung tissue of mice with acute paraquat poisoning, and discuss its pathological mechanism.Methods Fifty-eight healthy male mice were randomly divided into control group (n = 8) and poisoned group (n = 50). The mice in poisoned group were lavaged with 20% paraquat (50 mg/kg), and those in control group with equal amount of normal saline. The mice were sacrificed on the day of experiment in control group, and those in poisoned group at 6 hours and 1, 3, 7, 14 days after poisoning. The lung tissue was harvested to observe the changes in pathology of lung with hematoxylin and eosin (HE) staining. The positive expression of HO-1 was determined with immunohistochemistry, and the protein expression of HO-1 was determined with Western Blot. The contents of superoxide dismutase (SOD) and malonaldehyde (MDA) were determined.Results The mice showed shortness of breath and signs of exhaustion 1 hour after poisoning, getting worse on 3-5 days, but returned to normal 14 days after poisoning. Under the light microscope, it showed that the control group had no significant pathological changes in lung tissue. One day after the ingestion, pulmonary alveolar structure disorder, obvious hemorrhage, edema and infiltration of inflammatory cells were found. At 3 days, the pathological changes in the lung tissue were more pronounced. They were less pronounced on 7 days, and inflammatory changes disappeared on 14th day, but alveolar structure disorder remained. Immunohistochemical test showed that HO-1 was seldom expressed in the lung tissue, and a little amount was expressed in the mucosal epithelial cells of the airway in control group. It was shown that inflammatory cell and endothelial were mainly distributed in the mucosal epithelial cells of airway 1 day after poisoning followed by a gradually decrease tendence, and came to normal level of control group 7 days after poisoning. It was shown by Western Blot that HO-1 (gray value) in lung tissue increased 6 hours after poisoning (2.438±0.467 vs. 0.475±0.167,P< 0.01), peaked at 1 day (9.200±0.940 vs. 0.475±0.167,P< 0.01), continued to increase till 7 days after poisoning, and it lowered to normal level thereafter (0.825±0.260 vs. 0.475±0.167,P> 0.05). The SOD activity (μU/L) in lung tissue was lowered 6 hours after poisoning, and it was significantly lower than that of control group (649.681±13.951 vs. 1 167.051±15.744,P< 0.01), and it continued to decrease up to 14 days after poisoning (859.733±121.079 vs. 1 167.051±14.744,P< 0.01). MDA content (μmol/L) in the lung tissue homogenate was elevated 6 hours after poisoning with significant difference compared with that of the control group (4.542±0.266 vs. 3.705±0.176,P< 0.01). It peaked on day 1 (5.956±0.281 vs. 3.705±0.176,P< 0.01), then it declined and reached normal level 3 days after poisoning (4.134±0.168 vs. 3.705±0.176,P> 0.05).Conclusion HO-1 expression was increased significantly in lung tissue of mice with acute paraquat poisoning, which may be considered as an important protection mechanism against paraquat poisoning.

Background and purpose:The patients with aggressive lymphoma who have a poor prognosis and unlikely to be cured with conventional chemotherapy. This study was aimed to evaluate the effect of high-dose etoposide in mobilization followed auto-SCT in treating refractory lymphoma. Methods:40 patients [median age 33 (13-61) years] with refractory non-Hodgkin’s lymphoma (NHL, n=32) or Hodgkin’s lymphoma (HD, n=8) received high-dose etoposide [VP16 10-15 mg/(kg·d)×2 d] in mobilization in our center. Remission status prior to mobilization was PD (n=40). The use of such granulocyte colony-stimulating factor [G-CSF, 5-10μg/(kg·d)] mobilized peripheral blood stem cells (PBSC) after high-dose etoposide until the end of leukapheresis. Peripheral blood stem cell was collected and frozen in-80℃refrigerator. All these patients received auto peripheral blood stem cell transplantation (auto-PBSCT). Conditioning regimen was BEAM (n=19, 47.5%) or CBV (n=21, 52.5%). Results:Twenty-eight pa-tients (70%) were assessable for response after high-dose etoposide at a median pretreatment time of 39 days (range 17-172 days), 12 patients (30%) had no response. Median follow-up of 28 (4-66) months, 16 patients (40%) reached CR after auto-PBSCT. Fifteen of the 28 patients (53.6%) who had response to high-dose etoposide reached CR, 4 patients (14.3%) reached PR, 9 patients (32.1%) succumb to progression of disease. One of the 12 patients (8.3%) who had no response to high-dose etoposide reached CR, 1 patients (8.3%) reached PR, 10 patients (83.4%) succumb to progression of disease. The estimated 1-year OS and EFS were 69%and 56.7%respectively, 2-years OS and EFS were 63%and 52%respectively. The prognosis of the patients who had no response to etoposide was poor. The estimated 1-year OS and EFS were 25%and 16.7%respectively. Two group of comparison differences have statistics signiifcance (P<0.01). Conclusion: High-dose etoposide could be used in refractory lymphoma as rescue therapy in mobilization. It can increase the EFS and OS of patients who had response. The hematopoietic stem cells collection and hematopoietic reconstitution are not affected by etoposide.

BACKGROUND:Effects of different oxygen partial pressures on cytokine secretion of human adipose-derived stem cells have been differently reported. These differences may be caused by varying oxygen partial pressures.
OBJECTIVE:To investigate the influence of different oxygen partial pressures on cytokines secreted from human adipose-derived stem cells.
METHODS:Human adipose-derived stem cells were cultured in vitro and identified by its immunophenotype. Human adipose-derived stem cells were divided into five groups and cultured under different oxygen partial pressure conditions (1%, 3%, 5%, 10%, 21%) for 24 hours, respectively. With quantitative real-time PCR and enzyme linked immunosorbent assay, the secretion of cytokines, vascular endothelial growth factor, hepatocyte growth factor, nerve growth factor, keratinocyte growth factor, from human adipose-derived stem cells were analyzed on the gene and protein levels.
RESULTS AND CONCLUSION:Human adipose-derived stem cells were positive for CD71, CD73, CD90, CD105 and negative for CD34, CD45, CD54, HLA-DR. From the aspect of gene level, hypoxia (1%, 3%O 2 ) promoted the expression of vascular endothelial growth factor and nerve growth factor from human adipose-derived stem cells (P<0.01), and significantly elevated the expression of hepatocyte growth factor (P<0.05);however, there was no significant influence on keratinocyte growth factor under hypoxia (P>0.05). Based on the protein level, protein secretion of hepatocyte growth factor and vascular endothelial growth factor from human adipose-derived stem cells was increased under hypoxia (P<0.01), but no changes occurred in nerve growth factor and keratinocyte growth factor. After cultured under hypoxic environment, human adipose-derived stem cells were promoted to express gene vascular endothelial growth factor, hepatocyte growth factor and nerve growth factor, as wel as to secrete protein keratinocyte growth factor and vascular endothelial growth factor.

Objective To construct a geometric skull model by using three-dimensional reconstruction, computer tomography scanning and rapid prototyping technology and evaluate its significance in treatment of complex oral and maxillofacial deformities. Methods A cranial and a goldenhar syndrome patient with complex oral and maxillofacial deformities bone received continuous volumetric scanning of skull and the data acquisition was done by an electron computed tomography, by which reconstruction was performed and the obtained images saved as STL files. Then the data were input into rapid prototyping machine to make three-dimensional geometric model. Direct measurement, designation and surgery simulation could be done on this three-dimensional model. Then the mandibular mirror physical model was manufactured using rapid prototyping according to the normal side. Results A computer-aided model according to CT data could represent the three-dimensional anatomic structures and their relationships precisely. The replica exhibited dimensional errors ranging 0.02 mm to 0.53 mm. Which provided strong basis for accurate understanding of disease status and reasonable surgical plans and helped improve the curative effect of surgery. Conclusion Rapid prototyping can help surgeons in many ways for therapy of the complex oral and maxillofacial deformities.

objective To assess relationship between microvascular changes of hemodynamics of the fingertip and evolution of hypertension in patients with essential hypertension(EH)by enhanced power Doppler imaging(E-Flow).Methods The right ring fingertip'S blood flow peak systolic velocity(PSV),end-diastolic velocity(EDV),resistance index(RI)of finger ventral arteriole and nail bed arteriole in thirty-nine patients with EH were separately observed and were made contrast with 35 age-matched healthy subjects by E-Flow imaging.Results Compared with traditional colour Doppler flow imaging and power Doppler imaging,E-Flow imaging was clearer to demonstrate the distribution and movement of the fingertip'S microvascular.It was shown that the PSV and EDV in the ring finger ventral arteriole and nail bed arteriole of EH were decreased(P＜0.05),but the RI increased(P＜0.05),along with the evolution of EH.Conclusions Fingertip'S microvascular changes of hemodynamics could be a criterion to detect obstruction alternations of essential hypertension.E-Flow imaging iS superior to traditional color Doppler flow imaging and power Doppler imaging in differentiating microvascular and could provide a new method in assessing microvacular changes of patients with EH for clinic.

Objective　To study the diagnosis, treatment and prognosis of hepatolithiasis combined with cholangiocarcinoma. Methods　The clinical, pathological and follow-up data of 17 cases of hepatolithiasis combined with cholangiocarcinoma were retrospectively analysed. Results　The results showed that the incidence of cholangiocarcinoma in hepatolithiasis was 5% in this series. 17.6% of the patients were diagnosed as cholangiocarcinoma preoperatively. Tumor occurring in intrahepatic ducts was 88.2% and in hepatic porta ducts 11.8%. Nine cases were well-differentiated adenocarcinomas. Only 7(41.2%) cases were radically resected and their average survival time was 26.0 months. Eight(47.1%) patients underwent internal drainage with average survival time 12.4 months. 2(11.7%) cases subject to external drainage with survival time 3.6 months. Conclusions　If patients with hepatolithiasis have a long history of recurrent cholangitis, weight-loss in a short period, progressive jaundice or intractable abdominal pain, the possiblility of combined with cholangiocarcinoma should be considered. Resection of the tumor has a better prognosis than that of tumor unresected; and the prognosis of internal drainage is better than that of external drainage.