OBJECTIVETo study the influence of bone marrow mesenchymal stem cells (MSCs) transplantation on hypoxic pulmonary hypertension (HPH) in rats.

METHODSSD rats MSCs were separated, cultivated, identified and labeled by the green fluorescence protein (GFP) gene virus and transplanted in vitro. Healthy male SD rats were randomly divided into four groups: Normal control group (NC group) and HPH group (eight rats respectively), HPH+ MSCs transplantation group and HPH+ VEGF+ MSCs transplantation group (twenty-four respectively). The test employed atmospheric intermittent low oxygen method to establish the rat model of pulmonary hypertension and stem cells were transferred and transplanted. The rats' mean pulmonary artery pressure (mPAP) was observed; right ventricular hypertrophy index (RVHI) was calculated; the morphological change of lung small artery in various groups of rats was observed under the microscope; the distribution of lung small artery and adenovirus transfection fluorescently labeled MSCs was observed under a fluorescent microscope after 7, 14 and 28 days when stem cell was transplanted.

RESULTSFor NC group, the mPAP (mmHg) was 15.5 +/- 1.5 after twenty-eight days while the mPAPs for HPH , MSCs and MSCs+ VEGF were 26.1 +/- 1.9, 21.6 +/- 2.7 and 20.1 +/- 2.9 respectively which were apparently higher than that of NC group (P < 0.01) and compared with HPH group (P < 0.01), which declined clearly. There was no significant difference between MSCs and MSCs+ VEGF. After twenty-eight days, RVHI for NC group was 0.28 +/- 0.02 while the RVHI for HPH, MSCs and MSCs + VEGF were 0.43 +/- 0.07, 0.34 +/- 0.03 and 0.35 +/- 0.01 respectively which was apparently higher than that of NC group (P < 0.01) but which was clearly lower than that of MSCs and MSCs+ VEGF (P < 0.05) and there was no significant difference between MSCs and MSCs + VEGF. For HPH group, pulmonary arteriole wall became apparently thicker, the lumen became significantly narrow and nearly obstructed after twenty-eight days, the endothelial cells were incomplete; compared with HPH group, pulmonary arteriole wall of MSCs group became thin, the lumen was smooth and the completeness of endothelial cells was improved. Whereas for MSCs and MSCs + VEGF, these changes were not significantly clear.

CONCLUSIONAfter MSCs transplantation, mPAP and RVHI decline sharply and lung small artery remodeling is improved which partially reverses HPH process; there is no significant difference between VEGF together with MSCs transplantation group and pure MSCs.

Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; etiology ; metabolism ; surgery ; Hypoxia ; complications ; Male ; Mesenchymal Stem Cell Transplantation ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; pharmacology

Objective To explore the roles and possible mechanism of calcium-sensing receptor(CaSR) in cell cardiac hypertrophy model using angiotensin Ⅱ (Ang Ⅱ ).Methods The cultured neonatal rat ventricular myocytes were treated with Ang Ⅱ as cell cardiac hypertrophy model.Hypertrophic neonatal rat cardiomyocytes were treated with GdCl3(a specific agonist of CaSR) and/or with Ro318220(a specific inhibitor of PKC pathway).To evaluate the status of cardiac hypertrophy,cell diameter was observed by HE dyeing,and protein content was determined through coomassie brilliant blue protein kit.The intracellular calcium concentration( [ Ca2+]i) was determined by laser scanning confocal microscope.The protein expression of CaSR and PKC pathway were analyzed using Western blotting.Results ①Compared to the control group(0.1263 ± 0.0443),the protein expression of CaSR was increased in Ang Ⅱ group and in GdCl3 group(0.1963 ± 0.0375,0.2778 ± 0.0564,all P＜ 0.05).Moreover,compared with Ang Ⅱ alone,the increase was significant in GdCl3 group(P ＜ 0.05).②Compared to control group(222.70 ± 22.09),AngⅡ group(392.16 ± 36.85) remarkably increased [Ca2+]i(P＜ 0.05),and this increase of [Ca2+]i was further enhanced in GdCl3 group (502.60 ± 44.21) versus Ang Ⅱ group (P ＜ 0.05).③Compared to control group,Ang Ⅱ could induce cardiomyocyte hypertrophy,and GdCl3 enhanced the effect.Moreover,this enhancement was attenuated by Ro318220.④Compared to control group(0.27 ± 0.07,0.69 ± 0.06,0.87 ± 0.04),the protein expression of PKCα,PKCε and PKCδ was increased in Ang Ⅱ group(0.60 ± 0.16,1.02 ± 0.13,1.20 ± 0.18,all P＜ 0.05) and the protein expression of PKCα,PKCε was increased in GdCl3 group(0.82 ± 0.16,1.34 ± 0.12,all P ＜ 0.05).Moreover,compared with Ang Ⅱ group,the protein expression of PKCα,PKCε was obviously increased in GdCl3 group (all P ＜ 0.05);compared with GdCl3 group,the protein expression of PKCα,PKCε(0.41 ± 0.10,0.85 ± 0.14) was obviously decreased in Ro318220 group(all P ＜ 0.05).Conclusions CaSR is involved in cardiac hypertrophy induced by Ang Ⅱ through PKC pathway in cultured neonatal rat cardiomyocytes.

Objective To observe the curative effect of captopril and metoprolol in the treatment of chronic Keshan disease (CKD). Methods One hundred and ninty-five patients with CKD chosen from Juxian, Wulian, Yishui, Pingyi, Sishui and Zoucheng in Shandong Province were randomly assigned to control group, captopril group and metoprolol group according to NYHA cardiac functional grading. All cases were given diuretics, digitalis and vasodilating agents as routine treatment. On this basis, captopril and metoprolol was administered in captopril group and metoprolol group respectively. After 12 months of follow-up visit, the causes of cardiac death, hospitalization status and the changes of heart size, electrocardiogram, blood pressure and heart rate were all observed. Results It was found that the mortality of captopril group and metoprolo] group was 4.76% (3/63), 5.00% (3/60) respectively, both lower than the control group 10.61%(7/66). But this difference had no statistically significance(P=0.39). Besides, the hospitalization days of each year in captopril group and metoprolol group was respectively (19.12± 20.35) and(18.86±21.52)days, much more reduced than in the control group[(21.45±21.74)days, q=3.17, 3.38, P<0.05]. The detection rate of cardiothoracic ratio decreased in captopril group and metoprolol group [45% (27/60) and 40.4% (23/57)] After treatment showed more pronounced amelioration than the control group [18.6% (11/59), χ2=9.51,6.59, all P<0.0125], still the detection rate of cardiomegaly and invariability had no significant difference among three groups (χ2=2.50,4.75, all P>0.05). The elimination coefficient of ectopic rhythm in metoprolol group [56.5% (13/23)] was pronounced higher than the control group and captopril group [23.8% (5/21), 22.7% (5/22)], but differences had no statistically significance(P=0.0358,0.0331, all P>0.0125). Significant differences were found in systolic blood pressure(SBP), diastolic blood pressure(DBP) and heart rate(HR) in three groups prior and post-treatment(F=47.51,44.23,80.66, all P<0.01). The interaction of therapy and treatment time had influence on SBP and HR (F=3.19,37.44, all P<0.05), but had no influence on DBP(F=2.21, P> 0.05). There was no difference in SBP, DBP or HR among three groups before treatment(F=0.28,0.57,1.80, all P>0.05). After treatment, SBP and DBP in captopril group, metoprolol group and the control group[(109.0±10.9), (112.2±12.8), (114.7±13.2)mm Hg, (69.3±7.2), (72.1±9.5), (73.3±9.3)mm Hg] were all lowered compared with pre-treatment[ (117.1±13.4), (119.0±14.4), (117.6±14.1)mm Hg and (74.2±10.2), (76.3±10.8), (75.4±11.1)mm Hg, t=4.79,4.47,2.08,5.12, 4.32,2.15, all P<0.05]. HR was reduced in metoprolol group, being [(66.2±7.7), (75.9±11.5)times/min] before and after treatment(t=10.81, P<0.01), while it remained unchanged in captopril group and control group[(70.6±8.0), (72.6±10.5) times/min and (71.9±10.4), (73.8± 12.2)times/min, t=1.77,1.74, all P>0.05]. After treatment, both SBP and DBP of captopril group were significantly lower than that in the control group (q=3.52,3.56, all P<0.05); HR was reduced in metoprolol group, lower than that in captopril group and control group(q=5.44,3.73, all P<0.01). Conclusions Having a tendency of depressing mortality, captopril and metoprolol can reverse or delay myocardial remodeling and reduce admission rate in a safe,reliable and economic way, and are worth to be widely used in the treatment of chronic Keshan disease.

AlM:To observe the relationship between axial length and complications of phacoemulsification with intraocular lens ( lOL) implantation in high axial myopia eyes and normal axis eyes.
METHODS: A retrospective review of 843 consecutive patients ( 1 042 eyes ) of cataract extraction with phacoemulsification and lOL implantation in our hospital from February 2012 to February 2013 was performed. The patients were divided into two groups according to the axial length: 853 eyes were in normal axis group ( 21-24mm) and 189 eyes were in high axial myopia group (≥26mm). The two groups were compared regarding surgical complications, such as vitreous loss, posterior capsular rupture, nucleolus drop, and abnormal location of lOL.
RESULTS:Age was a risk factor in both groups. There was positive correlation between age and surgical complications, and between axial length and surgical complications, especially for complications with posterior capsular rupture and vitreous loss.
CONCLUSlON:As the results illustrate, in this survey, age and high axial lengthare statistically significant risk factors for incidence of complications of phacoemulsification. Anticipation of these complications and also preparation and prophylactic measures may decrease incidence of these complications.

BACKGROUNDEnamel decalcification in orthodontics is a concern for dentists and methods to remineralize these lesions are the focus of intense research. The aim of this study was to evaluate the remineralizing effect of casein phosphopeptide amorphous calcium phosphate (CPP-ACP) nanocomplexes on enamel decalcification in orthodontics.

METHODSTwenty orthodontic patients with decalcified enamel lesions during fixed orthodontic therapy were recruited to this study as test group and twenty orthodontic patients with the similar condition as control group. GC Tooth Mousse, the main component of which is CPP-ACP, was used by each patient of test group every night after tooth-brushing for six months. For control group, each patient was asked to brush teeth with toothpaste containing 1100 parts per million (ppm) of fluoride twice a day. Standardized intraoral images were taken for all patients and the extent of enamel decalcification was evaluated before and after treatment over this study period. Measurements were statistically compared by t test.

RESULTSAfter using CPP-ACP for six months, the enamel decalcification index (EDI) of all patients had decreased; the mean EDI before using CPP-ACP was 0.191 ± 0.025 and that after using CPP-ACP was 0.183 ± 0.023, the difference was significant (t = 5.169, P < 0.01). For control group, the mean EDI before treatment was 0.188 ± 0.037 and that after treatment was 0.187 ± 0.046, the difference was not significant (t = 1.711, P > 0.05).

CONCLUSIONCPP-ACP can effectively improve the demineralized enamel lesions during orthodontic treatment, so it has some remineralization potential for enamel decalcification in orthodontics.

Adolescent ; Caseins ; chemistry ; therapeutic use ; Dental Enamel ; drug effects ; Female ; Humans ; Male ; Orthodontics ; methods

Objective To explore the interaction between serum melatonin and the status of disease and probe the effect factor of serum melatonin change in asthmatic children.Methods Serum melatonin was measured in asthmatic children with 15 cases of mild persistent asthma,15 cases of moderate persistent asthma,15 cases of severe persistent asthma,15 cases of stable asthma and 15 cases of normal subjects by enzyme-linked immunosorbent assay(ELISA).Results The levels of serum melatonin in the 5 groups of mild persistent asthma,moderate presistent asthma,Severe Persistent asthma,Stable asthma,control subject were(22.76?5.16)ng/L,(16.79?3.35)ng/L,(11.54?1.45)ng/L,(22.06?3.36)ng/L,(28.72?4.32)ng/L,respectively.There were significant differences between any of them(Pa

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

AIM: To observe the effect of Ahmed glaucoma valve implantation intravitreal bevacizumab in the treatment of neovascular glaucoma ( NVG) .
METHODS:Twenty-two cases (22 eyes) who presented with NVG were first treated with intravitreal bevacizumab 0. 1mL ( 2. 5mg ), then with Ahmed glaucoma valve implantation after regression of iris neovessels. Cases were followed - up for 6 - 36 ( mean 24 ) mo with observation on visual acuity, IOP control, regression of iris neovessels, and complications during or after surgery.
RESULTS: Iris neovessels was regressed in different degree after injection within 1wk in 22 eyes. At final follow-up, the IOP of 18 eyes were all less than 21mmHg without any drugs and of 3 eyes with 1-3 kinds of anti-glaucoma drugs after combined Ahmed glaucoma valve implantation. The IOP of one eye was controlled after cryotherapy. The mean IOP dropped from 45. 36 ±8.13mmHg preoperatively to 15. 59 ± 3. 21mmHg postoperatively. IOP reduction was statistically significance between preoperative and postoperative ( P<0. 05) at final follow-up. Visual acuity was improved in 9 eyes (41%) and was no changed in 13 eyes. No serious complications were observed during or after intravitreous bevacizumab injection and Ahmed glaucoma valve implantation.
CONCLUSION: Ahmed glaucoma valve implantation and intravitreal bevacizumab in the treatment of NVG is useful and safe. It improves the success rate of surgery and preserves visual function, furthermore its complications are less.

Objective To investigate the effect and underlying mechanism of radioactive 125I seed implantation on the angiogenesis of transplanted human lung adenocarcinoma in nude mice.Methods An animal model of transplantd human lung adenocarcinoma was established by subcutaneous implanting A549 cells into nude mice.Twenty four tumor-bearing nude mice were randomly divided into 4 groups with different irradiation doses of blank control (without any treatment) and 0 MBq,22.2 MBq,29.6 MBq and by embedding radioactive 125I seeds with an 18 G implant needle.Tumor volumes were measured every 4 days until all mice were terminated 30 d later and the tumor growth curve was drawn.The microvessel density (MVD) in the tumor tissue was detected by immunohistochemistry S-P assay.The mRNA and protein levels of VEGF and HIF-1α of each group were detected by RT-PCR and Western blot,respectively.Results After embedding of 125I seeds,the tumor volumes of 22.2 MBq group (886 ± 97) and 29.6 MBq group (590 ± 107) were significantly smaller than those of control group (2 297 ± 149) at 54 d after administration (q =14.117,17.075,P ＜ 0.05),but there were no significant differences among 0 MBq group and control group,22.2 MBq and 29.6 MBq groups (P ＞ 0.05).The immunohistochemical CD34-positive staining demonstrated that MVD in 22.2 MBq group (522 ± 119) and 29.6 MBq group (491 ± 121) were decreased significantly compared with control group (922 ± 260) (q =4.826,5.197,P ＜0.05),but there were no significant differences among 0 MBq and control groups,22.2 MBq and 29.6 MBq groups(P ＞0.05).The mRNA expressions of VEGF and HIF-1α in 22.2 MBq group (0.279±0.0659,0.370 ±0.0857) and 29.6 MBq group (0.215 ±0.0620,0.278 ±0.0651) were significantly lower than those in the control group (q VEGFmRNA =18.881,17.211,q HIF-1αmRNA =15.376,14.733,P ＜0.05),but there were no significant differences among 0 MBq and control groups,22.2 MBq and 29.6 MBq groups(P ＞0.05).At the same time,the expression levels of VEGF and HIF-1α protein after 125I seed implantation were also obviously decreased in 22.2 MBq and 29.6 MBq groups (qvEGr =5.848,6.263,q HIF-1α =6.560,7.576,P ＜ 0.05),and no significant difference between 0 MBq and control groups(P ＞ 0.05) and between 22.2 MBq and 29.6 MBq groups (P ＞ 0.05).Conclusions Interstitial implantation with 125I seeds may potently inhibit angiogenesis in human lung adenocarcinoma xenografts of nude mice.

To obtain a number of extracellular penicillinase,the gene(penp)encoding penicillinse of Bacillus cereus ATCC10987 was cloned into expression vector in Bacillus subtilis,transformed into Bacillus subtilis DB104 deficient in two proteases.When recombinant bacteria was cultured in LB medium for 24 hours,the result of SDS-PAGE showed that the molecular weight of the protein was 28kDa and the penicillinase activity reached 339U/ml.By screening seven different fermentation media,the result showed that 4# medium is favorable to producing penicillinase by the recombinant cells than the others,with the yield of the enzyme being 1580U/ml.When the recombinant cells was cultured in 7 liter fermentor for 24h,the penicillinase activity reached 1255.8U/ml.