Objective:To provide reference for the treatment of lipoprotein glomerulopathy and to investigate the participation of clinical pharmacists in the whole treatment course. Methods: Three different treatment regimens, including immunosuppressive thera-py, dual plasma filtration therapy and lipid-lowering combined with reducing urinary protein therapy was respectively adopted for one patient with lipoprotein glomerulopathy, and the efficacy was evaluated. Clinical pharmacists assisted physicians in deciding treatment regimen, performed pharmaceutical care, adjusted medication and analyzed the prognosis of the patient during the follow-up. Results:1. Immunosuppressive therapy was ineffective for the patient;2. The dual filtration acted quickly, while the expense was high and the disease was easy to relapse;3. The third therapy was relatively safer and more economical with long-term effect. Conclusion:Combi-nation therapy of lowering lipid and reducing urinary protein is the most suitable treatment regimen for the patient with lipoprotein glo-merulopathy. Clinical pharmacists play an important role in the whole course of treatment to assist physicians in obtaining the maximum benefit of patients.

This study aimed to investigate the effects of Shiquan Dabu Tang (SDT) on growth and angiogenesis of subcutaneously implanted tumors, hepatic metastases, and incision-implanted tumors after surgical removal of primary colon tumor in mice.

Objective To predict the secondary structure and the B cell epitopes of human heparanase protein, and to identify its immunogenicity. Methods The flexible regions of secondary structure and the B cell epitopes of human heparanase amino acid sequence were predicted by DNAStar and Bcepred software. The multiple antigenic peptides (MAP) of the epitopes were synthesized in 8-branch form. Rabbits were immunized with the 8-branch MAPs mixed with a universal T-helper epitope human IL-1β peptide (VQGEESNDK, amino acid 163-171 ). The immunogenicity of the synthesized peptides was evaluated by ELISA, Western blot and immunohistochemistry. Results Amino acid 1 -15 ( MAP1), 279-293 (MAP2) and 175-189(MAP3) of large-subunit of human heparanase protein was predicted as the most potential epitopes of human heparanase protein. All the three synthesized MAPs induced high titer of antibodies. ELISA, Western blot and immunohistochemistry analysis showed all the three MAPs could produce high titer serum antibodies, antibodies induced by MAP1 and MAP2 had high specific binding activity , and MAP2 antibody showed the strongest binding activity with liver cancer tissues. Conclusion The large-subunit No. 1-15, 279-293 amino acid of human heparanase protein may be the B cell preponderant epitopes and the strongest immunogenicity may be No. 279-293 peptide, which provided a theoretic basis for the antibody and vaccine development of heparanase subunit peptide.

AIM: To evaluate the blood compatibility of a new bioartificial reactor membranous material (propylene-acidamide grafted poly propylene membrane, PP-g-AAm) in vitro. METHODS: Contacted PP-g-AAm membrane and PP (polypropylene) memb rane with platelet-rich plasma in a swing bed, 37 ℃, to simulate the conditions in vivo, and another group of PRP without any membranes was set as control group. ELISA was used to study the expression of ?-thromboglobulin, and flow cy tometry was used to study CD62P and CD63 expressio n of the activated blood platelets after contacting the two kinds of membranes w ith PRP. Scanning electrical microscopy was used to study the configuration and numbers of platelet cells adhered on the membranes. RESULTS: After contacting with PRP 30 min, ?-TG expression show ed marked difference between the two kinds of material groups and the control gr oup (P

Objective To compare the clinical effect of neoadjuvant chemoradiotherapy (nCRT) and neoadjuvant chemotherapy (nCT) in the treatment of locally advanced esophageal squamous cell carcinoma.Methods The retrospective cohort study was conducted.The clinicopathological data of 156 patients with local advanced esophageal squamous cell carcinoma who were admitted to the Zhongshan Hospital of Fudan University from January 1,2010 to December 31,2015 were collected.Among 156 patients,59 undergoing nCRT were allocated into the nCRT group and 97 undergoing nCT were allocated into the nCT group.Patients in the nCRT group and nCT group respectively received 2 cycles chemotherapy by the TP regimen+40 Gy radiotherapy (2 Gy/d) and 2 cycles chemotherapy by the TP regimen.Patients were evaluated by imaging examinations after 6 weeks neoadjuvant therapy completion,and then underwent abdominal and right chest-left cervico three-incision thoracoscopic surgery.Observation indicators:(1) treatment situations;(2) postoperative pathological examination;(3) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed once every 3 months within 2 years and once every 6 months after 3 years up to January 2017.Follow-up included levels of tumor markers [carcinoembryonic antigen (CEA) and SCC-Ag],thoracic or abdominal computed tomography (CT),neck and abdominal ultrasonography and gastroscopy or PET/CT examination if necessary.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range) and comparison between groups was analyzed using the nonparametric test.Count data were analyzed using the chi-square test or Fisher exact probability.Comparison of ordinal data was done by the nonparametric test.The survival rate was calculated using the life table method and survival was analyzed by the Log-rank test.Results (1) Treatment situations:all the patients in the 2 groups were able to burden neoadjuvant therapy and thoracic esophagectomy.Six patients in the nCRT group and 15 in the nCT group had conversion to open surgery.Operation time,volume of intraoperative blood loss,cases with postoperative readmission of ICU,cases with complications,cases with perioperative death and duration of hospital stay were (201 ± 25) minutes,(137± 66)mL,5,24 (10 with pulmonary complications,8 with anastomotic leakage,3 with hoarseness,2 with cardiovascular complications and 1 with chylopleura),0,12 days (range,9-93 days) in the nCRT group and (195±20) minutes,(133±58) mL,8,30 (11 with anastomotic leakage,10 with pulmonmy complications,4 with hoarseness,2 with cardiovascular complications,1 with postoperative hemorrhage,1 with delayed gastric emptying and 1 with chylopleura),1,11 days (range,9-78 days) in the nCT group,respectively,with no statistically significant difference between the 2 groups (x2 =0.883,t =0.102,0.692,x2 =0.048,1.541,Z =0.225,P＞ 0.05).(2) Postoperative pathological examination:R0 resection rate was 96.6％ in the nCRT group and 93.8％ in the nCT group,with no statistically significant difference between the 2 groups (x2 =0.589,P＞0.05).Results of postoperative pathological examination showed that G0,G1,G2 and G3 of tumor regression grade were respectively detected in 18,16,7,18 patients in the nCRT group and 4,5,4,84 patients in the nCT group,with a statistically significant difference between the 2 groups (Z=-7.151,P＜0.05).Stage 0,Ⅰ,Ⅱ,ⅢA,Ⅲ B and ⅣA of postoperative ypTNM stage were respectively detected in 16,9,23,4,6,1 patients in the nCRT group and 4,9,37,6,34,7 in the nCT group,with a statistically significant difference between the 2 groups (Z=-4.890,P＜0.05).The down-staging was detected in 48 patients of the nCRT group and 50 patients of the nCT group,with a statistically significant difference between the 2 groups (x2=13.957,P＜0.05).(3) Follow-up and survival situations:of 156 patients,153 were followed up for 12-82 months,with a median time of 36 months.The 1-,3-,5-year overall survival rates were 88.1％,61.4％,34.9％ in the nCRT group and 81.4％,43.8％,23.1％ in the nCT group,with a statistically significant difference between the 2 groups (x2=4.336,P＜0.05).Conclusion The nCRT in the treatment of locally advanced esophageal squamous cell carcinoma can enhance postoperative pathological response rate,down-staging rate and overall survival rate compared with nCT,without increasing incidence of perioperative complications.

Objective To evaluate the effects of optimal pharmacotherapy according to guideline on treating chronic heart failure(CHF)in real world clinical practice. Methods A total of 231 consecutive outpatients with reduced left ventricular ejection fraction(LVEF≤40%)and enlarged left ventricular end diastolic diameter(male ＞55 mm, female ＞60 mm)were recruited from January 2001 to June 2009. All patients were treated with optimal pharmacotherapy according to guideline recommendations and followed up to December 31,2009. Mortality, rehospitalization and changes of heart size and cardiac function at baseline and at the end of follow-up period were analyzed. Results(1)14 patients were lost during follow-up (6. 1%), and follow-up was complete in 217 patients(93.9%). 97.2% and 98.2% patients were prescribed angiotensin converting enzyme(ACE)inhibitors and β-blockers(βB). Combined of ACE inhibitors and BB use was applied in 95.3% patients. The target dose of ACE inhibitors and βB were reached in 50. 7% and 37.3% patients.(2)Lower mortality and re-hospitalization rates were observed in this cohort: all-cause morality, average annual mortality was 11.5% and 3.9% respectively. Rehospitalization rate was 27.6%.(3)Left ventricular end-diastolic diameter(LVEDD)decreased from (68.2 ±7.2)mm to(62. 2 ±9. 6)mm. LVEDD value was normal or near normal(male≤60 mm, female ≤55 mm)in 43.2% patients. LVEF improved form(29. 8 ±7. 5)% to(43. 3 ± 11.8)%, LVEF was ＞40% in 60.4% patients, LVEF was ≤ 40% but increased ≥ 10% after treatment in 22.9%patients. Conclusion Optimal pharmacotherapy according to guideline can improve prognosis of outpatients with CHF.

Objective To explore the serum levels oi macrophage migration inhibitory factor (MIF),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in children with mycoplasma pneumoniae pneumonia (MPP) and their clinical significance.Methods The serum levels of MIF,TNF-α and IL-6 were detected by enzyme-linked immuno sorbent assay in 42 children with acute MPP (acute MPP group),26 children with recovery MPP (recovery MPP group),25 children with bacterial pneumonia (bacterial pneumonia group),and 30 normal healthy children (healthy control group).Correlations between MIF and TNF-α as well as IL-6 were analyzed in MPP group.Results The serum levels of MIF,TNF-α and IL-6 in acute MPP group were significantly higher than those of recovery MPP group,bacterial pneumonia group and healthy control group (all P ＜ 0.01,0.05),whereas there was no significant difference between recovery MPP group and healthy control group (P ＞ 0.05).Serum levels of MIF,TNF-α and IL-6 in bacterial pneumonia group were significantly increased compared with those in healthy control group (all P ＜ 0.05).There were positive correlations between MIF and TNF-α as well as IL-6 in acute MPP (r =0.76,0.82,all P ＜ 0.05),whereas there was no correlation between MIF and TNF-α as well as IL-6 in recovery MPP(r =0.26,0.31,all P ＞ 0.05).Conclusions MIF,TNF-α and IL-6 may play significant roles in the pathogenesis of MPP,and combined detection of the three factors has great referential value in assessing the severity and predicting the prognosis for MPP.

Objective To build a foundation for determination of C reaction protein,C reaction protein was expressed and purified,and the immune reactivity of the purified protein was identified.Methods The CRP cDNA was amplified by RT-PCR from human liver cDNA library and inserted into expression vector pCRTT/NT.The recombined plasmid CRP-pCRTT/NT which expressed the fusion protein of CRP was then transferred into lysogenic host strain E coli.BL21 (DE3).The target protein was identified using SDS- polyacrylamide gel electrophoresis (SDS-PAGE).Affinity chromatography was used for protein purification.The immune reactivity of purified CRP was identified by Western blot using anti-CRP specific antibody.Results Recombiant human CRP was expressed in inclusion bodies of E.coli with a six histamine tag.The purify of recombinant protein was detected by SDS-PAGE as a single band at 30 000 and was identified by Western blot.Conclusions A plasmid expressed CRP protein is constructed and the purification system of CRP protein is established.The immune reactivity of the purified protein is identified by Western blot,which makes a good base for the preparation of CRP test kit.

Objective To explore the effect of blocking BTLA-HVEM (herpesvirus entry mediator-B and T lymphocyte attenuator) pathway on dendritic cell function and the related immunological mechanisms. Methods Murine BTLA extracellular domain eukaryotic expression vector psBTLA was constructed by gene recombination and transfected CHO by Lipofection method. Mouse bone marrow cells were induced to differentiate into DCs by GM-CSF plus IL-4. Expression of BTLA and HVEM on DCs was detected after HSPT0-TC-1 peptide complex stimulation by FACS. Expression of BT-1 and secretion of IL-12 were detected after HSP70-TC-1 peptide complex plus psBTLA transfected CHO culture supernatant stimulation on DCs. Pretreated DCs co-cultured with the same genetic background mouse splenocytes and lymphocytes proliferation and cytokine secretion were detected. Effect of psBTLA gene transfer in vivo on BT-1 expression of DCs and tumor growth on tumor-bearing mice was detected. Results Extracellular domain of murine BTLA was successfully constructed, psBTLA stable transfection CHO cells were obtained and expression of BTLA extracellular domain(sBTLA) was detected the in its culture supernatant. BTLA and HVEM expression of DCs were increased after stimulation by the antigen peptide complex. When DCs were treated with antigen peptide complex plus culture supernatant containing sBTLA, B7-1 expression and IL-12 secretion were increased. Co-cultured with splenocytes, lymphocytes proliferation and cytokine secretion, such as IL-2 and IFN-γ,, were also increased. Gene transfection with psBTLA in vivo promoted B7-1 expression on DCs and inhibited cervical cancer cells growth. Conclusion Blockade of BTLA-HVEM inhibitory pathway with sBTLA can further improve DCs function, activation of lymphocytes and promote antitumor immune response.

Objective To investigate the synergistic therapy effects of B and T lymphocyte attenuator(BTLA) extracellular domain in combination with heat shock protein 70 (HSP70)-TC-1 antigen peptide complex on the mouse model of cervical cancer and the related immunological mechanisms. Methods(1)Detecting the BTLA and herpesvirus entry mediator (HVEM) gene expression in the tumor microenvironment after C57BL/6 mice were inoculated with TC-1 tumor cells by realtime PCR; BTLA,HVEM expression on tumor infiltrating lymphocytes cell surface were detected by flow cytometry (fluorescence intensity). (2) According to different treatments, tumor-bearing mice were divided into 5 groups, which was injected with pcDNA3. 1 (empty vector plasmid as control), psBTLA (vector plasmid which expresses BTLA extracellular domain), HSP70 (HSP70-TC-1 cell peptide complex), HSP70 +pcDNA3.1 or HSP70 + psBTLA, respectively. The weight of tumor was recorded. The expression of immunoregulatory genes in tumor microenvironment were detected. The change of lymphocyte amount and cytotoxicity were detected too; lymphocyte proliferation activity was measured by tritium thymidine incorporation assay; the concentration of interleukin (IL) 2 and interferon-γ(IFN-γ) in supernatants of spleen lymphocyte were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) BTLA gene expression was gradually increased after tumor cells inoculation. The highest expression level was 2. 83 + 0. 35 at 14th day, which had statistical significance difference with the 7th day expression of 1.66±0. 25 (P < 0. 05). While HVEM mRNA expression did not change significantly (P > 0. 05). The 7th and 14th day after TC-1 cells inoculation, the average fluorescence intensity of BTLA expression on the surface of tumor infiltrating lymphocytes was 33.5 and 51.8, respectively, in which there was statistically significant difference (P <0. 05); while the difference of HVEM expression was not statistically significant (57. 2 vs 49. 3 ,P >0. 05). (2)The 28th day after inoculation, tumor inhibition rate of HSP70 + psBTLA group was 88%, which was significantly higher than other treatment groups (P <0. 05). The 28th day after TC-1 cells inoculation, combination therapy not only promoted IFN-γ and IL-2 gene (3. 12 + 0.71,3.20 + 0. 62)expression but also reduced transforming growth factor-β (TGF-β), Foxp3 and IL-10 expression (0. 25±0. 03,0. 19 +0. 03,0. 31 +0. 04;P <0. 05). It also promoted CD8+ T lymphocyte infiltration(52 +6)/high power field, cytotoxicity (65.5±2.4) %, proliferation (15.0 × 103 cpm) and cytokine IL-2 , IFN-γsecretion(824±51), (1096±112) pg/ml, which were all significantly higher than other groups (P <0. 05). Conclusion The effect of immunotherapy on tumor can be augmented by the combination of psBTLA which expresses extracellular domain of BTLA and HSP70-TC-1 tumor antigen peptide complex,which could improve the expression of the related immunoregulatory genes to establish a much better microenvironment in favor of anti-tumor immune response against the mice model of the cervix carcinoma.