Objective To assess the feasibility of using biodegradable poly (lactic-co-glycolic acid) (PLGA) scaffold with controlled release of rhBMP-2 and its effect on the osteogenic activity of MSCs in vitro. Methods The PLGA scaffolds with rhBMP-2 sustained release system were made by combination of porogen-leaching and freeze-drying. The drug release kinetics was measured by enzyme-linked immunosorbent assay (ELISA) in vitro. MSCs were isolated from the bone marrow of human and cultured for 3 passages. Then, MSCs were seeded onto PLGA scaffolds. PLGA scaffolds without rhBMP-2 were used as experimental controls . Scanning electron microscopy(SEM) was used to observe the morphology of MSCs. The cell proliferation was determined by MTT assay. Results The rhBMP-2 was encapsulated into PLGA scaffolds and it was found to be continuously released from the scaffolds. The scaffolds have evident enhancing effect on ectopia osteogenesis. Conclusion PLGA scaffold containing rhBMP-2 is a new promising tissue engineering scaffold.

BACKGROUND: It is proposed that elevated serum homocysteine is an important independent risk factor for ischemic stroke (IS), and 5, 10-methylenetetrahydrofolate reductase (MTHFR) is the key enzyme for homocysteine metabolism. The relationship between genetic mutation of MTHFR and IS remains controversial.OBJECTIVE: To examine the association of hyperhomocysteinemia and two MTHFR gene polymorphisms with IS in Northwest Chinese population.DESIGN: Case-control study.SETTING: Department of Neurology, First Hospital Affiliated to Jilin University, and Department of Neurology, Xijing Hospital, Fourth Military Medical University of Chinese PLA.PARTICIPANTS: Ninety-seven consecutive patients with ischemic stroke (71 males and 26 females) treated between November 2001 and May 2002were recruited, who were diagnosed by CT scan or MRI in the Department of Neurology, Xijing Hospital, Fourth Military Medical University of Chinese PLA. The control group consisted of 94 subjects (58 males and 36 females) without history of ischemic stroke. All the subjects were free of intracranial hemorrhage, cancer, renal dysfunction, and none used multivitamins or estrogen.METHODS: Serum homocysteine was measured by fluorescence polarization immunoassay. Polymerase chain reaction-restriction length polymorphism (PCR-RFLP) method was employed to detect the genotype at the two sites of C677T and A1298C in MTHFR gene.MAIN OUTCOME MEASURES: Serum homocysteine levels and the genotypic frequency frequencies of the two mutations of MTHFR.RESULTS: The 677T allele frequency was 59.3% in IS patients and 44.7% in the controls, showing significant differences (P=0.006), but no difference in 1298C allele frequency was detected between the two groups (22.7% vs 19.7%, P ＞ 0.05). Homozygous 677TT genotype was closely associated with hyperhomocysteinemie (P ＜ 0.01). In multivariate logistic regression analysis,677T gene mutation and hyperhomocysteinemie were all associated with the IS, with an OR of 1.870 and 1.031 (P＜ 0.05), respectively.CONCLUSION: Hyperhomocysteinemie is a risk factor of IS, and C677T mutation significantly increases homocysteine levels, and serves also as an independent genetic risk factor of IS.

BACKGROUND: Multiple sclerosis(MS) is a chronic autoimmune disease induced by the interaction between genetic and environmental factors. Its pathogen and the mechanism of the relapse and remission m the course of the disease are still unknown. Most of the MS research centers are looking for the pathogenic polypeptide epitope in proteolipid protein(PLP), myelin sheath basic protein (MBP) and oligodendrocyte glycoprotein (MOG) OBJECTIVE: To compare the proliferation of T cell lines(TCL) in MS induced by myelin sheath and delipidated myelin sheath towards 11 components of myelin sheath to mainly search the possible pathogenic polypeptide epitope in PLP, and investigate the possible effects of abnormal dcgrease in myelin sheath.DESIGN: A case-controlled trial.SETTING: Department of neurology in a hospital of a university.PARTICIPANTS: Mononuclear cells(MNC) of 16 MS cases(clinical relapsing-remitting type, patients did not receive any immunosuppresant for at least 3 months when their peripheral blood samples were taken) and 12 HLA-DR15 healthy volunteers were furnished by Dr. Trotter JL of MS Research Center of Washington University from the cell database.INTERVENTIONS: MS-TCL and normal TCL were induced twice by stimulation with myelin sheath and delipidated myelin sheath in vitro by cell culture in vitro. TCL proliferation was tested by 11 antigens including PLP,MBP, M87-106, P30-49, P40-60, P89-106, P95-117, P117-137,P139-151, P178-191, and P185-206.MAIN OUTCOME MEASURES: Difference of scintillation counting in every minute of every well, and the stimulative index of each well were calculated, and the mean wells with positive proliferation of TCL towards each antigen were confirmed as well.RESULTS: The general specific proliferation towards myelin sheath antigens was bigger in MS group than control group 5.49 ±5.31 to 3.10 ± 3. 17, and delipidated myelin sheath-induced TCL was bigger than myelin sheath-induced one 5. 49 ± 5.31 to 3.41 ± 4. 83 . Delipidated myelin sheath significantly changed the immune responses of MS group,especially the changes of responses towards P30-49, P40-60, P89-106,P117-137, P139-161, and P185-206 were significant compared with that the control group only responded to two polypeptides, which indicated that the antigen epitope of MBP, PLP, M87-106, P95-117, P40-60, and P185-206 might have significance in the triggering of MS autoimmune responses.CONCLUSION: TCL induced by MS myelin sheath has different proliferation towards antigen components of myelin sheath from control group. Delipidated myelin sheath significantly increases TCL proliferation in MS group, which suggests that if MS patients developed abnormal degrease in myelin sheath, TCL would produce autoimmune response towards self-myelin sheath, MBP, PLP and its polypeptide segments all can trigger MS or aggravate the state of the illness. Our finding supports the hypothesis of MS autoimmune pathogenic mechanism.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Fingers ; Humans ; Male ; Middle Aged ; Neuralgia ; therapy ; Young Adult

Objective To study the pharmacokinetics and pharmacodynamics of the 40/60 premixed recombinant human insulin injection preparation,and to compare with 30/70 preparation,regular insulin,and neutral protamine Hagedorn (NPH).Methods In this positive control,single dose,open label,Latin square crossover study,20 male healthy volunteers were recruited from May 2006 to March 2007,and divided into four groups.On 4 test days,40/60 preparation,30/70 preparation,regular insulin,and NPH were administered to each of the 4 groups,the interval was 7-70 days before 2 test days.The pharmacokinetics and pharmacodynamics were evaluated by euglycemic glucose clamp technique.Results According to the analysis of variance,there were statistically significant differences in pharmacokinetics and pharmacodynamics of the 4 insulin formulations between the 4 groups (all P ＜ 0.05).For the 40/60 premixed recombinant human insulin,the pharmacokinetic parameter time to peak (Tmax) and mean retention time (MRT) were (105.00 ±24.33) minutes and (321.77 ± 56.29) minutes,respectively;the glucose-lowering effects reflected by the pharmacodynamic parameter Tmax and MRT were (167.75 ± 26.48) minutes and (248.33 ± 14.96) minutes,respectively.Compared with 30/70 premixed recombinant human insulin,40/60 preparation showed no significant differences in the pharmacokinetics parameters of blood insulin concentration,including peak concentration [(91.67 ± 13.03) mU/L vs.(84.96 ± 14.75) mU/L,P =0.119],Tmax [(105.00 ± 24.33) minutes vs.(122.25 ± 39.35) minutes,P =0.128],MRT [(321.77 ± 56.29) minutes vs.(332.12 ± 49.20) minutes,P =0.645] and area under the curve in 0-16 hours [AUCIns 0-16,(24 918 ± 6 610)h · mU/L vs.(26 768 ± 8 032)h· mU/L,P=0.084];however,statistically significant differences were observed in AUCIns0-4 [(16 991 ± 3 673) h · mU/L vs.(12 407 ± 3 441) h · mU/L,P =0.042] and AUCIns 0-8 [(23 283 ± 4 939) h · mU/L vs.(19 397 ±5 314)h · mU/L,P =0.046].Pharmacodynamic parameters showed no statistically significant differences (all P ＞ 0.05).Compared with 30/70 premixed insulin,the relative bioavailability of 40/60 premixed insulin was (118.9 ± 35.9) ％,and the relative biological effectiveness was (106.6 ± 35.2) ％.There was no clinically significant abnormalities in the safety indexes before and after the tests.No hypoglycemic events,allergic reactions,or local injection adverse reaction occurred in this trial.Conclusions The 40/60 premixed recombinant human insulin preparation demonstrated different properties in insulin absorption in 8 hours after injection compared with the 30/70 preparation,mainly because of the difference in proportions of short-and intermediate-acting insulin in the mixture.This new premixed insulin may provide a new option for personalized diabetes management.

Objective:To study the efficacy of stereotactic radiotherapy (SRT) combined with gemcitabine plus cisplatin (GP)-based chemotherapy on local recurrent non-small-cell lung cancer (NSCLC) after radiotherapy. Methods:Thirty-eight patients with local recurrent NSCLC were treated with SRT(3.00-4.85 Gy per fraction, five times every week, totally 8-12 fractions, total course of 2-3 weeks). Before SRT the patients were given two cycles of GP-based chemotherapy (gemcitabine 1 000 mg/m~2 d 1 and d 8;cisplatin 30 mg/m~2 d 1-3) and at the end of SRT they were given 4 cycles of GP-based chemotherapy. Results:Of the 38 patients,7 patients had complete response, and 23 patients achieved partial response. The overall immediate response rate was 78.95% (30/38). The 6-month and 12-month total survival rates were 71.05% and 10.53%,respectively. The median survival time was 7.8 months. Conclusion:SRT combined with GP-based chemotherapy had better short-term efficacy and the patients were tolerable. No severe short-term radiation-induced injury was observed. The long-term radiation-induced injury and long-term efficacy need further investigation.

Objective To study the pharmacokinetics and pharmacodynamics of recombinant human insulin preparations SciLin TM R and Humulin (R) R,and to evaluate their bioequivalence in Chinese healthy volunteers.Methods In this positive control,single dose,open label,randomized cross-over study,20 male healthy volunteers were recruited from March to October 2007,and tested on two experimental days with an interval of 7-14 days.The volunteers were divided into two groups with a random number table,one group was injected with SciLin TMR for the first time and Humulin (R) R for the second time,the other group was injected with the opposite.The pharmacokinetics and pharmacodynamic properties were evaluated by euglycemic glucose clamp study.Results Time to peak concentration [Tmax,(105.8 ± 19.1) minutes vs.(103.5 ± 18.1) minutes,P =0.389) and time to maximum glucose infusion rate [TGIRmax,(132.8 ± 16.8) minutes vs.(132.8 ± 18.6) minutes,P =0.697] for SciLin TMR and Humulin(R) R were similar.The relative bioavailability of SciLin TMR was (102.2 ± 7.6) ％,and the relative biological effectiveness was (107.4 ± 18.8) ％.The 90％ confidence interval(CI) of peak concentration(Cmax) and area under the curve of blood glucose concentration at 0-10 hours (AUCIns 0-10) of SciLin TM R were 99.32 ％-102.62 ％ (equivalent range 70％-143 ％) and 98.98 ％-104.99 ％ (equivalent range 80％-125％),respectively;90％ CI of the maximum glucose infusion rate (GIRmax) and AUCGIR0-10 were 97.36％ ～ 103.49％ (equivalent range 70％-143％) and 98.72％-113.54％ (equivalent range 80％-125％),respectively,indicating that SciLin TMR and Humulin (R) R was bioequivalent.There was no clinically significant abnormalities in the safety indexes before and after the tests.During the trial,no hypoglycemic events,allergic reactions,or local injection adverse reaction occurred.Conclusion The studied recombinant human insulin preparation SciLin TMR may be bioequivalent as Humulin (R) R.

Objective To study the characteristics of pharmacokinetics and pharmacodynamics of insulin dry powder inhalation and its relative bioavailability as compared with subcutaneous injection of regular insulin. Methods In this open,single-center,randomized,two-period,cross-over,euglycemic glucose clamp study,18 healthy volunteers(14 men and 4 women),aged(24.9?1.7)years,with body mass index(20.6?1.2)kg/m~2, received the insulin dry powder inhalatin(80 U)or regular insulin(15 U)subcutaneous administration.The blood samples of this study at 0,20,30,40,50,60,70,80,90,100,110,120,135,150,165,180,195, 210,225,240,270,300,330,360,390,420,450 and 480 rain were taken for serum insulin measurement, meanwhile,glucose infusion rates(GIR)were determined per 5 minutes over a period of 8 hours.Results The C_(max)were(57.9?17.8 vs 114.5?29.7)mU/L(tested vs reference preparation),T_(max)were(46.7?45.6 vs 107.8?33.7)min,GIR_(max)were(3.35?0.98 vs 5.17?1.75)mg?kg~(-1)?min~(-1)and T_(GIRmax)were(88.3?17.0 vs 151.9?34.6)min.The relative bioavailability was(10.26?2.25)%,and the relative bioefficacy was(14.33?7.26)%.Conclusion The study shows that insulin dry powder inhalation is absorbed via lungs and its action sets in earlier than that of the regular insulin injected subcutaneously.These pharmacokinetie and pharmacodynamic data may provide a reliabe guide for further clinical trial.

BackgroundThe morphological examination of macular disease is very important information for the early diagnosis.Spectral domain optical coherence tomography (SD-OCT) is a gold criteria in the diagnosis of macular diseases,but its clinical application is limited in the eyes with refractive medium opacity.The resolution ratio of conventional B-scan sonography is unsatisfactory.20 MHz B-scan sonography may be a new approach to maculopathy.ObjectiveThis study was to evaluate the clinical value of 20 MHz B-scan ultrasound by comparing morphological changes of different maculopathies observed by 20 MHz ultrasound and spectral-domain OCT.MethodsA prospective study was designed.51 eyes of 51 patients with macular diseases who were determined by Tianjin Medical University Eye Center were enrolled in this study.Spectral-domain OCT was used to image the macular area,and then the 20 MHz B-scan ultrasound was used to examine the same eye to assess the value of 20 MHz B-scan ultrasound diagnosis.The macular diseases were typed according to the OCT manifestation.The characteristics of 20 MHz ultrasound for the same maculopathy were analyzed.All the patients subscribe the informed consent before the study.ResultsIn 10 eyes with macular hole determined by spectral-domain OCT,shallow dome-shaped lesion with central hollow was exhibited on the ultrasound image.The macular edema,retinal neuroepithelium layer detachment and retinoschisis were found in 16 eyes,6 eyes and 4 eyes by spectral-domain OCT,but the 20 M Hz B-Scan ultrasound presented with the focal belt-shaped elevation with non-echo zone below.In contrast,in 15 eyes of retinal pigment epithelium detachment in OCT,spectral-domain OCT showed stronger and thicker belt-shaped echo.Conclusions20 MHz B-scan ultrasound has certain value in the diagnosis and differentiation for several common maculopathies.Although it is not accurate as OCT in recognizing diseases,it could play an important role in refractive media opacity eye.

Objective To investigate the effect of continuous blood purification (CBP) on pro-or anti-inflammatory immune function and prognosis in patients with sepsis. Methods One hundred and two patients with sepsis were randomized into the CBP group (60 cases) and the control group (42 cases). The patients in the control group were treated with conventional therapy, and the patients in the CBP group received at least 72 h CBP treatment additionally. The APACHEⅡ score, SOFA score, the 28 day survival rate and ICU length of stay were recorded and levels of spleen Th1, Th2 were assessed by FACS flow cytometry. Enzyme linked immunoadsordent assay (ELISA) was used to determine the levels of serum IL-1, IL-10 and TNF-α before and at 24, 48, 72 h after the treatment. Results The APACHEⅡscore and SOFA score decreased markedly in the CBP group after the treatment (Р <0.05). The period of staying in ICU of patients in the CBP group was shorter than that of patients in the control group (Р < 0.05). There was no significant difference of the 28 day survival rate between the two groups (91.6% vs 71.2%, Р > 0.05). Compared with the control group, levels of IL-1, IL-10 and TNF-α were decreased markedly, and the ratio of Th1 / Th2 was increased significantly at 72 h after the treatment in the CBP group (Р < 0.05). Conclusion CBP can eliminate inflammatory mediators, and help to enhance the immune function, and restore the balance of Th1 / Th2 in patients with severe sepsis.