The latest developments of techniques that prolong the half-life of protein drugs were reviewed.The therapeutic potential of several proteins is limited by their rapid clearance from the body,resulting in the need for frequent treatment of individuals to maintain drug levels.Several techniques including chemical modification,protein fusion,microsphere,glycosylation,and protease-resistant variants,and so on,prevent rapid degradation of such drugs and prolong their half-lives in vivo.Several successful protein drugs with prolonged half-life have already been on the market or on clinical trials.The principles,efficiencies and scopes of application of these techniques were described,compared and discussed.

OBJECTIVE:To establish an RP-HPLC method for the determination of doxorubicin hydrochloride concentration in human plasma.METHODS:Tianhe Kromasil C18 was used as column,A solution of methenol-0.01 mol?L-1ammonium dihydrogen phosphate-acetic acid(30200.1) was used as the mobile phase at a flow rate of 1.0 mL?min-1.The detection of wavelength was set at 233 nm.The sample size was 20?L and the column temperature was 35℃.The internal standard was daunorubicin.RESULTS:The calibration curve was linear in the range from 0.25～20.16 ?g?mL-1(r=0.999 6);the average methodological recovery was 96.45%(RSD=2.16%).CONCLUSIONS:The method is rapid,accurate,reproducible and easy to use in clinical detection of blood concentration.

OBJECTIVE:To establish a RP-HPLC method to determinate the plasma concentration of paclitaxel in tumor patients.METHODS:Paclitaxel was extracted from plasma with organic phase(ethyl-acetate)by two-step extraction on Tianhe Kromasil C18 column(250 mm?4.6 mm,5 ?m)with a mobile phase consisted of acetonitrile-methanol-water(40∶25∶40)at a flow rate of 1 mL?min-1.The detective wavelength was set at 227 nm and the column temperature was maintained at 35 ℃.RESULTS:The linear range of paclitaxel was 0.05～5.00 mg?L-1(r=0.999 7)with average recovery rate at 98.75%～100.44%.Both intra-day RSD and inter-day RSD were less than 5%(n=5).The plasma concentration-time profile in 11 patients after iv infusion of paclitaxel was in line with a two-compartment model.CONCLUSION:This established method is simple,accurate,reproducible and applicable for clinical determination of blood drug concentration and pharmacokinetic studies.

Objective: To study the relationship between the individual psychological characteristic of medical students and the way parents bring up them. Methods: Data on SCL-90, 16PF,EMBU of 1550 medical students were analysed. Results: It is showed that girl students have experienced more of the parents' emotion, understanding and favouritism, while the boy students have experienced more of their parents' refusal, deny, punishment, severity and excessive interference. There are more only-one-children who experience parents' emotion, understanding, interference and excessive protection. Conclusions: Parents' bad breeding attitude and behavior influence children's psychological health and forming of the individual characteristic directly or indirectly

A 68?year?old female patient was admitted to the hospital for multiple masses in the mouth and lungs as well as on dorsal hands for more than 20 days without obvious subjective symptoms. No abnormalities were found by physical examination. Dermatological examination showed two bean?sized dark?red nodules on the upper jaw as well as one pigeon egg?sized dark?red nodule on the left dorsal hand, and all the nodules were hard with smooth surfaces and limited mobility. Positron emission tomography?computed tomography (PetCT) revealed multiple metastases to the brain, lymph nodes, lungs, gastrointestinal tract, both kidneys, multiple bones and intermuscular tissues. Pathology of nodules from the upper jaw showed lowly differentiated tumor cells with osteoid matrix, chondroid structures and tumor bone in local areas, and immunohistochemical examination of tumor cells found positive staining for S100(focally), vimentin, CD99, P63 and Ki?67(60%), but negative staining for keratin. A diagnosis of osteosarcoma of the right side of the upper jaw was considered. Pathology of nodules from the dorsal hand revealed no obvious abnormalities in the epidermis, while there was a diffuse infiltration of medium?to large?sized histiocyte?like cells in the whole dermis with cell atypia and irregularly red?stained bone matrix and tumor bone in some regions. Immunopathology showed positive staining for Ki67(60%), and negative staining for CD3, CD10, CD20, Bcl?2, and Bcl?6. A diagnosis of cutaneous metastasis of osteosarcoma was made. The patient refused further treatment and died 6 months after the onset of lesions.

Objective To investigate the regulatory effect of CsA on the expression of NK cell inhibitory receptor ILT4 and cytotoxicity of NK cells.Methods NK cells treated with CsA ( 10 mg/L) or DMSO for 12,24 and 36 h were chosen as three experimental groups and control groups respectively.RTqPCR and flow cytometry were performed to detect the alteration of ILT4 at the mRNA and protein level respectively.The expression of HLA-G in human gastric cancer cell line BGC-823 and human placental choriocarcinoma cell line JEG-3 were measured at the same time,and then the cytolytic activity of the untreated NK cells and NK cells treated with CsA for 36 h against BGC-823 and JEG-3 cells was determined with MTT.One-way analysis of variance was employed to compare the different ILT4 expression at different time points after medication; Dunnett test was performed to carry out the pairwise comparison between each mean.The difference of HLA-G expression between JEG-3 cells and BGC-823 cells,and the difference of NK cell cytolytic activity against JEG-3 cells and BGC-823 cells were analyzed by student's t-test.Results RT-qPCR assay indicated that the relative levels of ILT4 mRNA in NK cells treated with CsA for 12,24 and 36 h in turn were 0.99 ± 0.27,1.79 ± 0.29,6.79 ± 0.64,and those of their contrast groups treated with DMSO were 0.86 ±0.11,0.94 ±0.12,1.06 ±0.17.The expression of ILT4 in NK cells treated with CsA for 24 h or 36 h was higher than that in NK cells of their contrast groups respectively ( t value of 4.69,14.99,P ＜0.05,respectively),but there was no significant difference between the two groups of NK cells treated for 12 h ( t =0.78,P ＞0.05 ).Through flow cytometry,the positive rates of ILT4 protein expression in NK cells treated with CsA for 12,24 and 36 h [(5.16 ± 0.42 ) ％,( 6.23 ± 0.48 ) ％,( 23.8 ± 1.5 ) ％]were higher than those in NK cells after treatment with DMSO for 12,24 and 36 h respectively[(3.08 ±0.19)％,(3.35 ±0.12)％,(3.36 ±0.21 )％ ;t value of 7.70,10.06,20.72,P＜0.01,respectively].The expression of ILT4 in NK cells treated for 36 h was much higher than that in NK cells for 12 and 24 h at the mRNA and protein level (t value of 16.38,14.12 ;21.81,20.56,P ＜ 0.01,respectively).Meanwhile the killing rates of NK cells treated with 10∶1 effector-target ratio CsA on BGC-823 cells (low HLA-G expression) were ( 8 1.96 ± 2.80 ) ％ ( before treatment) and ( 60.23 ± 1.57 ) ％ ( after treatment),which were higher than those on JEG-3 cells (HLA-G-overexpression) [(53.46 ±2.21 )％ ( before treatment),(28.30 ± 1.85 ) ％ ( after treatment)].The changes of cytotoxicity of NK cells treated with CsA against target cells showed that CsA inhibited the killing activity of NK cells to BGC-823 and JEG-3 cells (t value of 11.74,15.16,P＜0.01,respectively),and the inhibitory rates were (26.48 ±2.42)％ and (47.10 ±1.59 ) ％ respectively.CsA had a higher killing rate inhibition on JEG-3 than on BGC-823 ( t =12.31,P ＜0.01 ).Conclusion CsA induces upregulation of ILT4 in NK cells,and the cytotoxicity of NK cells to tumor cells can be affected by interaction of ILT4 and HLA-G.

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Cell Differentiation ; Cell Proliferation ; Drug Resistance, Neoplasm ; Glioma ; pathology ; Glycoproteins ; metabolism ; Humans ; Neoplastic Stem Cells ; metabolism ; pathology ; physiology ; Neovascularization, Pathologic ; etiology ; pathology ; Peptides ; metabolism ; Radiation Tolerance

Abstract LRR-RLKs plays an important role in multiple signal transduction in plant.One full-length cDNA encoding a LRR-RLKs homologue was isolated from maize through in silico cloning and named as ZmLRRPK1(GenBank accession:EU873320).The predicted ZmLRRPK1 protein has 594 amino acids with an estimated molecular mass of 66kDa and an isoelectric point of 5.42.ZmLRRPK1 has the typical domain of LRR-RLKs.RT-PCR analysis indicated that ZmLRRPK1 expression was induced by ABA,mannitol and salt in coleoptiles of maize and kept high level during 24 hours.

Objective To compare the prescrotal orchiopexy and traditional inguinal orchiopexy in the clinical treatment of children with low cryptorchidism.Methods Seventy-two patients(78 testes)who underwent orchiopexy in our hospital during March 2006 to May 2011 were retrospectively analyzed.And the undescended testis could be manipulated beyond the external inguinal ring under anaesthesia.Matching conditions were age differences among 3 months,same preoperative testicular positioning,same surgeon and same side.Using the paired study of 1 to 1,all the patients were divided into 2 groups: prescrotal orchiopexy(group A)and the traditional inguinal orchiopexy(group B),each group included 36 patients(39testes).Mean age was 5.4 years(group A)and 5.5 years(group B).The time of operation and restore standing,success rate and complications,including hernia,hydrocele,testicular atrophy and ascent were compared between the 2 groups.Results All the patients were successfully operated.The average surgical time for the prescrotal and inguinal groups were 33 and 41 min(P =0.0022),and average time of standing was 1.2 and 5.4 d(P =0.0003).All the patients had no wound infection.Followup ranged from 3 to 65 months.No hernia,hydrocele,testicular atrophy and ascent were identified in either group.The ratios of successful surgery were 100％ in the two groups.Conclusions The prescrotal orchiopexy is simple,safe,and effective in the cases that testis could be pushed down through the external inguinal ring.Compared with traditional inguinal approach,the advantages of prescrotal approach are shorter operative time,fewer traumas,less pain,faster recovery and cosmetic results.

Objective To investigate the effects of lipopolysaccharide(LPS) on myelin phagocytosis during Wallerian degeneration after early peripheral nerve injury in rats.Methods Fifty male Wistar rats were recruited and randomly divided into LPS group(n=20),model group(n=20) and sham group(n=10).The right sciatic nerves of rats in the LPS and model groups were cut and sutured end-to-end,while the sciatic nerve of sham group rats were only exposed.Immediately after surgery,the rats in LPS group were given microinjections of LPS(2 g/L) into the surgical site in a final volume of 1 μL,and the rats in other two groups were injected with the same volume of saline.The sciatic nerves were taken at 1.5 h,24 h and 7d after surgery.Real-time quantitative PCR(qRT-PCR) was applied to detect the dynamic expressions of IL-1β mRNA and MCP-1 mRNA.Immunofluorescence staining was used to test the expression of CD68+ macrophages in sciatic nerves.HE staining was used to observe the pathological alterations of sciatic nerves tissue.ORO staining was used to observe sciatic nerves demyelination.LFB staining was used to detect the sciatic nerves myelin.Sciatic function index was used to evaluate the recovery of motor function in rats.Results Compared with the model group,qRT-PCR indicated that the expression of IL-1β and MCP-1 from LPS group were increased at 1.5 h and 24 h after surgery(P<0.001, P<0.001),respectively.Compared with the model group,the expression of CD68+ cells was increased significantly at 7th day after surgery(P<0.05).Histological examination showed that compared with the model group,a lot of inflammatory cells and Schwann cells were found at sciatic nerve stump in the LPS group at 7th day after operation.ORO staining showed that the degree of demyelination in the LPS group was higher than that in the model group.LFB staining showed that the sciatic nerve stump demyelination appeared in both model group and the LPS group at 7th day after operation,but compared with the model group,myelin debris clearance in the LPS group was significantly accelerated(P<0.05).Finally,compared with the model group,the SFI in the LPS group was increased significantly at 20 d after surgery(P<0.05).Conclusions The results confirm that LPS is possible to manipulate the innate immune response to accelerate myelin clearance during Wallerian degeneration after early peripheral nerve injury in rats.