1.Current Status and Progress on the Techniques for Prolonging Half-life of Protein Pharmaceuticals
Lei BIAN ; Yi-Feng SHI ;
China Biotechnology 2006;0(02):-
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.
2.Determination of Doxorubicin Hydrochloride Concentration in Human Plasma by RP-HPLC
China Pharmacy 2001;0(08):-
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(30200.1) 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.
3.Determination of Plasma Concentration of Paclitaxel in Tumor Patients by RP-HPLC
China Pharmacy 2005;0(15):-
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.
4.Updates on study of glioma stem cells.
Zhi-hua ZHOU ; Liang YI ; Xiu-wu BIAN
Chinese Journal of Pathology 2007;36(3):201-203
AC133 Antigen
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Animals
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Antigens, CD
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metabolism
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Cell Differentiation
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Cell Proliferation
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Drug Resistance, Neoplasm
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Glioma
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pathology
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Glycoproteins
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metabolism
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Humans
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Neoplastic Stem Cells
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metabolism
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pathology
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physiology
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Neovascularization, Pathologic
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etiology
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pathology
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Peptides
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metabolism
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Radiation Tolerance
5.Regulatory effect of CsA on the expression of NK cell inhibitory receptor ILT4 and cytotoxicity of NK cells
Yuanquan SI ; Yi ZHANG ; Xiaokun BIAN ; Nan LU ; Yanfei JIA
Chinese Journal of Laboratory Medicine 2011;34(11):1029-1033
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.
6.A case of cutaneous metastasis of osteosarcoma
Junwei FAN ; Yi BIAN ; Jincheng DONG ; Wei HOU ; Xuefeng WAN
Chinese Journal of Dermatology 2016;49(7):482-484
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.
7.Study on the Relationship Between the Individual Psychological Characteristic of Medical Students and the Way Parents Bring up them
Ling WANG ; Zhihong GONG ; Peiling BIAN ; Jingbo XIE ; Yi ZHANG
Chinese Journal of Medical Education Research 2002;0(01):-
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
8.cDNA Cloning of Maize Protein Kinase Gene ZmLRRPK1 and Its Expression Analysis
Ming-Di BIAN ; Chuan DENG ; Zhong-Yi BAI ;
China Biotechnology 2006;0(07):-
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.
9.Comparison of single prescrotal incision and traditional inguinal orchiopexy
Qinjun YI ; Dawei HE ; Zedong BIAN ; Tao LIN ; Xuliang LI ; Junhong LIU ; Feng LIU ; Yi HUA
Chinese Journal of Urology 2013;(2):120-122
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.
10.Mechanisms of cordycepin on improving renal interstitial fibrosis via regulating eIF2α/TGF-β/Smad signaling pathway.
Liu-bao GU ; Rong-wen BIAN ; Yue TU ; Hao HU ; Yi-gang WAN ; Wei SUN
China Journal of Chinese Materia Medica 2014;39(21):4096-4101
OBJECTIVETo investigate the effects and mechanisms of cordycepin,an effective component of cordyceps militaris, on renal interstitial fibrosis (RIF) and its related eIF2α/TGF-β/Smad signaling pathway.
METHODFirstly, 15 C57BL/6 mice were randomly divided into 3 groups,the control group (Group A), the model group (Group B) and the cordycepin-treated group (Group C). After renal interstitial fibrotic model was successfully established by unilateral ureteral obstruction (UUO), the mice in Group C were intraperitoneally administrated with cordycepin(5 mg x kg(-1) d(-1)) and the ones in Group A and B were administrated with physiological saline for 5 days. At the end of the study, the obstructed kidneys were collected and detected for the pathological changes of RIF, and the mRNA expressions of collagen type I (Col I) and α-smooth muscle actin (α-SMA) in the kidney by Northern blot. Secondly, after renal tubular epithelial (NRK-52E) cells cultured in vitro were exposed to transforming growth factor (TGF) -β with or without cordycepin, the mRNA expressions of Col I and collagen type IV( Col IV) by Northern blot, and the protein expressions of eukaryotic initiation factor 2α (eIF2α), phosphorylated eIF2α ( p-eIF2α), Smad2/3 and phosphorylated Smad2/3 (p-Smad2/3) were tested by Western blot.
RESULTIn vivo, cordycepin alleviated RIF in model mice, including improving fibrotic pathological characteristics and mRNA expressions of Col I and α-SMA. In vitro, cordycepin induced the high expression of p-elF2α, and inhibited the expressions of p-Smad2/3, Col I and Col IV induced by TGF-β in NRK-52E cells.
CONCLUSIONCordycepin attenuates RIF in vivo and in vitro, probably by inducing the phosphorylation of eIF2α, suppressing the expression of p-Smad2/3, a key signaling molecule in TGF-β/Smad signaling pathway, and reducing the expressions of collagens and α-SMA in the kidney.
Actins ; analysis ; Animals ; Deoxyadenosines ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Protein-Serine-Threonine Kinases ; physiology ; Signal Transduction ; drug effects ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; antagonists & inhibitors ; physiology