Objective: In order to solve the problem of ginkgolide B (GB) for preparing the intravenous injection dosage form, such as it’s low solubility, rapid elimination in vivo and poor stability in long-term storage, a novel biodegradable polysaccharide polymer was used as a carrier to encapsulate GB into nanoparticles with sustained-release profile, which then was freeze dried for keep its stability. Methods: GB nanoparticles (GB-NP) were prepared by coacervation method with hydrophilicity polymer. The Design-Expert 8.0 software was applied for experimental design. The formulation was finalized by investigating the concentration of GB, the mass ratio of GB to polymer, and the pH of the polymer solution with size and polydispersity indexas the evaluation parameters. The optimum formulation and technique were selected by response surface method. Then the nanoparticle suspension was further freeze dried and in vitro release behavior was tested in PBS containing 30% ethanol. Results: The optimum prescription conditions were as follows: the concentration of GB was 1.5 mg/mL, the ratio of GB mass to polymer mass was 0.1 and the polymer solution pH was set at 5.0. The entrapment efficiency was (99.64 ± 0.45)% with the drug loading ratio of (9.04 ± 0.04)%. Average particle size of GB-NP was (192.8 ± 2.8) nm with a preferable PDI of 0.18 ± 0.03. The freeze-drying conditions were described as follows: using 1% mannitol as freeze- drying supporting agent, the solution of GB-NP was pre-frozen for 12 h at −80 ℃ and dried for 24 h at −40 ℃. The release behavior of free GB was rapid with the cumulative release rate reaching (64.74 ± 3.95)% during the first hour, while the cumulative release rate of GB in nanoparticles was (36.90 ± 1.41)% during the first hour. Conclusion: The GB-NP using this special biodegradable polysaccharide polymer can improve GB’s dissolution behavior in water and make GB sustained release in vitro, which is beneficial for preparing GB’s intravenous injection dosage form.

As the first national“preventive treatment of disease”preventive healthcare service pilot, Changning District in Shanghai has carried out the construction of TCM preventive healthcare service platform under the guidance of district leaders from 2009. By introducing the construction process and implementation key points of the pilot linkage patten of medical institution and social healthcare institutions in Changning District, this article put forward the construction ideas of “Platform for Regional Traditional Chinese Medicine Preventive Healthcare Service”, and further proposed challenges and opportunities for deepening reforms.

Objective To study the radiographic features of the chordoid glioma and its differential diagnosis,and increase neuroradiologists'awareness of this newly described tumor,facilitating prospective diagnosis.Methods Three patients with chordoid glioma were reviewed retrospectively to determine whether any characteristic radiographic findings and clinical-pathologic findings would emerge,and the relevant literatures were reviewed.Routine CT(2 cases)and MR(3 cases),contrast-enhanced MR(3 cases)were performed.Results The masses were well circumscribed and located in the region of the hypothalamus/third ventricle-suprasellar region(2 cases)and intrasuprasellar region(1 case).Tumors were isointense to gray matter on CT scans and were isointense or hypointense on T_1WI and hyperintense on T_2WI.In two patients,vasogenic edema extended into the optic tracts and cystic or necrotic area was seen.All cases were remarkably enhanced following contrast administration.Conclusions Radiologic findings of the chordoid glioma has some unique features.Tumor,in the region of the hypothalamus-third ventricle-suprasellar region,if it is well circumscribed and remarkably enhancing following contrast administration,should be included in the differential diagnosis.

Objective To express in prokaryotic system and to analyze the antigenic specificity of EB virus(EBV) latent membrane protein 2(LMP2) multi-epitopes gene rich of T cell and B cell epitopes.Methods Using on-line prediction service, T cell epitopes and B cell epitopes of EB virus latent membrane protein 2 were predicted. The genes rich of CTL and th cell epitopes were selected as the candidate gene sequences, while B cell epitopes around them were taken into account. The finial selected multi-epitope gene was synthesized after being optimized according to prokaryotic codon bias and inserted into prokaryotic expression vector pET32a( + ) to get the recombinant plasmid: pET32a( + )/EBV-LMP2 multi-epitopes. After transformed into E. coli BL21 (DE3) and induced by IPTG, the target multi-epitopes gene can be expressed as Trx-His fusion protein. The expression products can be identified by SDS-PAGE and Western blot. Moreover, rabbit serum antibody to EBV membrane protein and nasopharyngeal carcinoma(NPC) patient serum were used respectively to detect the antigenic specificity of the multi-epitopes. Meanwhile, 6-8 weeks female BALB/c mice were immunized with EBV-LMP2 multi-epitope at 2 week intervals, three times in all, Trx-His protein and PBS were set as the control groups. At the second week after the last immunization, the mice were sacrificed. LDH and indirect ELISA were taken to detect the specific spleen CTL activityand specific IgG in serum, which reflected the immunogenicity of the EBV-LMP2 multi-epitope. Results Two amino acid sequences which locate at the LMP2 (aa195 -232 ) and LMP2 (aa419-436 ) were selected and connected in series to be the target gene. The recombinant plasmid containing EBV-LMP2 multi-epitope gene successfully constructed and the target protein was expressed in E. coli BL21 ( DE3 ). The relative molecular mass(Mr) of The expression products is about 27 × 103 , which matches up to the expected Mr. The antigenic specificity of the multi-epitopes protein was identified by Western blot and the multi-epitopes protein also can be detected by rabbit serum antibody to EBV membrane protein and NPC patient serum respectively. In the result of the animal experiment, EBV-LMP2 multi-epitope was able to induce the specific CTL activity in BALB/c mice. With the increasing of the effector: target ( E: T) 1: 5,1: 10, 1: 25, the CTL activity was also increased wih( 12.52% + 2.59% ), (21.80% + 1.08% ), (23.68% + 3.74% ) respectively; EBV-LMP2 multi-epitope was able to induce LMP2-specific antibody response(A490 =0.258 +0.040) as compared with Trx-His protein(A490 =0.095 +0.011) and PBS(A490 =0.068 +0.014,P＜0.05＝. Conclusion The EBV-LMP2 multi-epitopes gene was designed successfully and expressed precisely in prokaryotic expression system with good antigenicity and immunogenicity.

OBJECTIVETo observe the effect of Jianpi Bushen Qingchang Huashi Recipe (JBQHR) on proliferation and migration of bone marrow mesenchymal stem cells (BMSCs).

METHODSBMSCs were isolated and cultured in vitro with adherence screening method to prepare cell suspension. No drug intervention was given to BMSCs in the vehicle control group. JBQHR at 0.39, 0.78, 1.56 µg/mL was added in BMSCs of low, mid, and high dose JBQHR groups for co-incubation. Its effect on the proliferation of BMSCs was detected by CCK-8. BMSCs migration and chemotactic ability was detected using Transwell method. Each dose JBQHR combined ERK kinase inhibitor U0126 was set up as control. The phosphorylation of extracellular regulated protein kinase (ERK) and CAMP responsive element-binding protein (CREB) were detected by Western blot.

RESULTSCompared with the vehicle control group, the proliferation of BMSCs and BMSCs migration number could be promoted in the 3 JBQHR groups (P < 0.05). Besides, the proliferation of BMSCs was better in mid and high dose JBQHR groups than in the low dose JBQHR group (P < 0.05). Compared with the vehicle control group, the phosphorylation of ERK and CREB could be elevated in the 3 JBQHR groups (P < 0.05), and could be inhibited by U0126 (P < 0.01). Compared with the low dose JBQHR group, the phosphorylation of ERK increased in mid and high dose JBQHR groups with statistical difference (P < 0.05).

CONCLUSIONJBQHR could promote the proliferation and migration of BMSCs, and its mechanism might be related to ERK/CREB signaling pathway

Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; MAP Kinase Signaling System ; Mesenchymal Stromal Cells ; cytology ; drug effects

Objective To evaluate the nucleated red blood cell (NRBC)count of Sysmex XN-9000 automatic hematology anal-ysis lines comparing with manual method,and verify the accuracy of the analyzer results.Methods 60 blood samples with more than 1% of NRBCs detected by XN-9000 were counted NRBCs by traditional manual microscopy in blood smears,and verified the analyzer results.Results According to the reliability analysis,the results of total 60 samples were all within the range of the reliability;correlation analysis showed that correlation coefficient (r)of group NRBC (%)1~10 and>10 were 0.972 1 and 0.996 2,respectively.There were no significant differences between them (P>0.05).Conclusion Compared with manual method,the results of NRBC count of XN-9000 were within the range of reliability,and showed good correla-tion.The analyzer test results of NRBC were accurate and reliable and could be applied to the detection of clinical samples.

The study on tumor metabolism has been gradually be-come a hot spot in recent years .A lot of proteins involved in the regulation of tumor metabolism especially the glucose transporter protein 1(GLUT1).As a key regulatory factor mediating energy metabolism within tumor cells , GLUT1 can regulate the glucose intake and maintain the basic level of metabolism in tumor cells . More importantly, the abnormal expression of GLUT1 was asso-ciated with many kinds of tumors , of which GLUT1 was used to meet the energy requirement for the fast growth of tumor .Thus GLUT1 also played a crucial role in growth , differentiation and metastasis of tumor cells and prognosis of tumors .Meanwhile , as three-dimensional crystal structure of GLUT 1 was determined , it is possible to design the small molecular inhibitors of GLUT 1, which can realize “starve to death” tumor cells.GLUT1 can be a particularly attractive target for tumor treatment and interfer-ence.The relationship between abnormal expression of GLUT 1 protein and tumor metabolism was reviewed . Moreover , the mechanism of tumor metabolism regulated by GLUT 1 protein ex-pression and treatment of cancers were discussed , which may provide references for future research and clinical treatment .

Objective To investigate the incidence and risk factors of surgical site infection ( SSI ) in patients with colorectal cancer .Methods Clinical data of patients with colorectal cancer undergoing surgical treatment in Jiaxing First Municipal People’ s Hospital from October 2011 to December 2014 were retrospectively reviewed.The gender, age, underlying diseases, smoking history, preventive medication, abdominal surgery history , type of surgery , preoperative levels of hemoglobin and albumin , use of laparoscopy, use of stapler, combined organ resection, TNM staging, American Society of Anesthesiologists ( ASA) score was documented .Multivariate logistic regression analysis was performed to identify the risk factors of SSI .Results A total of 773 patients were enrolled in the study , and SSI was observed in 144 cases (18.63%).Multivariate logistic regression analysis showed that use of laparoscopy ( OR =0.35, 95%CI:0.15-0.79,P <0.05), use of stapler (OR =0.59, 95% CI: 0.39-0.88,P <0.05) were protective factors for SSI, while diabetes (OR=2.11, 95% CI: 1.25-3.58,P<0.01), liver cirrhosis (OR=2.12,95%CI:1.18-3.79,P<0.05), ASA score (3-4 points) (OR=2.01,95%CI:1.20-3.58, P<0.01), combined organ resection (OR=2.17,95% CI:1.20-3.92,P<0.05), and anastomotic leak (OR=6.85, 95%CI:3.01-15.63,P<0.01) were risk factors for SSI.Conclusions The incidence of SSI is high in patients with colorectal cancer undergoing surgery .Use of laparoscopy and stapler may reduce the incidence of SSI .

AIM:To investigate the safety and effect of femtosecond laser-assisted cataract surgery with Cionni modified capsular tension ring (MCTR) implantation in the management of traumatic lens subluxation.METHODS: Totally 11 patients (11 eyes) with traumatic lens subluxation were divided into three groups according to the severity of lens dislocation, ranging from 90° to 120° (4 eyes), 120° to 180° (5 eyes) and 180° to 270° (2 eyes).The contact LenSx femtosecond laser cataract surgery platform was applied to create the capsulotomy, prepare nuclear fragmentation and make corneal wound creation.Anterior vitrectomy was performed in some patients during the surgery.After capsular retractors insertion and phacoemulsification, the MCTR was inserted to the capsular bag and fixed to the sclera.Finally, the IOL was implanted into the capsular bag.Postoperative visual acuity, intra-and post-operative complications, anterior capsular opening, IOL and MCTR position and intraocular pressure (IOP) were assessed.RESULTS:The duration of follow-up was 2mo.All the operations were completed successfully.Five eyes underwent cataract surgery combined with anterior vitrectomy.Four eyes had been inserted with 2-eyelet MCTR and seven eyes with 1-eyelet MCTR.The best corrected visual acuity (BCVA) after operation was better than 0.5 in 4 eyes, between 0.3 and 0.5 in 3 eyes, between 0.1 and 0.3 in 3 eyes, and less than 0.1 in 1 eye.Compared with preoperative BCVA, the difference was statistically significant (P<0.05).All the IOLs were stably centered and the eyelet of MCTR was fixated steadily between the iris and the anterior capsule.The common intra-and post-operative complications were subconjunctival hemorrhage, incomplete capsulotomy, residual cortex, secondary glaucoma and posterior capsular opacification.CONCLUSION:Femtosecond laser-assisted cataract surgery can improve the success rate of capsulorhexis, and reduce the difficulty of nuclear fragmentation.Femtosecond laser-assisted cataract surgery combined with MCTR implantation is an ideal surgical method for traumatic lens subluxation.

In order to investigate the expression of nerve growth factor (NGF) and hypoxia inducible factor-1α (HIF-1α) and its correlation with angiogenesis in non-small cell lung cancer (NSCLC), paraffin-embedded tissue blocks from 20 patients with NSCLC were examined. Twenty corresponding para-cancerous lung tissue specimens were obtained to serve as a control. The expression of NGF, HIF-1α, and vascular endothelial growth factor (VEGF) in the NSCLC tissues was detected by using immunohistochemistry. The microvascular density (MVD) was determined by CD31 staining. The results showed that the expression levels of NGF, HIF-1α and VEGF in the NSCLC tissues were remarkably higher than those in the para-cancerous lung tissues (P<0.05). There was significant difference in the MVD between the NSCLC tissues (9.19±1.43) and para-cancerous lung tissues (2.23±1.19) (P<0.05). There were positive correlations between NGF and VEGF, between HIF-1α and VEGF, and between NGF and HIF-1α in NSCLC tissues, with the spearman correlation coefficient being 0.588, 0.519 and 0.588, respectively. In NSCLC tissues, the MVD had a positive correlation with the three factors (P<0.05). Theses results suggest that NGF and HIF-1α are synergically involved in the angiogenesis of NSCLC.