Objectives: To investigate the effects of adhesion and proliferation of bone mesenchymal stem cells (BMSCs) in the surface of lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol PLGA-[ASP-PEG] tri-block polymer scaffolds, try to find a new biomaterial to induce seed cells in vitro for bone tissue engineering. Methods: Modified PLGA with polyethylene glycol (PEG) and asparagic acid (ASP) that has many ligands, and synthesis PLGA-[ASP-PEG] polymer material. BMSCs were cultured in PLGA-[ASP-PEG] polymer material and PLGA used as control group. Through precipitation method, MTT assay and total cellular protein detection to test the adhersion and proliferation of BMSCs. Scanning electron microscope is used to observe cells appearance. Results: BMSCs on the surface of PLGA-[ASP-PEG] polymer scaffolds are adherention to the culture flask, the number of cells is much higher than PLGA’s. The precipitation method suggest that adhesion and proliferation of BMSCs on the surface of PLGA-[ASP-PEG] is much higher than the control group(P

OBJECTIVETo investigate the effect of graded hypothermia on neuropathologic alterations of neonatal rat brain after exposed to hypoxic-ischemic insult at 37°C, 33°C, 31°C, and 28°C, respectively, and to observe the effect of hypothermia on 72-kDa heat shock protein (HSP72) expression after hypoxic-ischemic insult.

METHODSSeven days old Wistar rats were subjected to unilateral common carotid artery ligation followed by exposure to hypoxia in 8% oxygen for 2 hours at 37°C, 33°C, 31°C, and 28°C, respectively. The brain temperature was monitored indirectly by inserting a mini-thermocouple probe into the temporal muscle during hypoxia. After hypoxia-ischemia their mortality was assessed. Neuronal damage was assessed with HE staining 72 hours after hypoxia. HSP72 expression at 0.5, 24, and 72 hours of recovery was immunohistochemically assessed using a monoclonal antibody to HSP72.

RESULTSHypoxia-ischemia caused 10.5% (2/19) of mortality in rat of 37°C group, but no death occurred in 33°C, 31°C or 28°C groups. HE staining showed neuropathologic damage was extensive in rats exposed to hypoxia-ischemia at 37°C (more than 80.0%). The incidence of severe brain damage was significantly decreased in 33°C (53.3%) and 31°C groups (44.4%), and no histologic injury was seen in the 28°C group of rats. Expression of HSP72 was manifest and persistent in the rat brain of 37°C group, but minimum in the rat brain of 28°C group.

CONCLUSIONMild and moderate hypothermia might prevent cerebral visible neuropathologic damage associated with hypoxic-ischemic injury by decreasing stress response.

Animals ; Animals, Newborn ; Body Temperature ; Female ; HSP72 Heat-Shock Proteins ; metabolism ; Hypothermia ; Hypoxia-Ischemia, Brain ; pathology ; Pregnancy ; Rats ; Rats, Wistar

OBJECTIVETo study sedative, analgesic, antidepressant and anti-inflammatory effects of the extracts from Piper boehmeriaefolium var. tonkinense.

METHODEtOAc soluble constituents and EtOH soluble constituents were administered 50, 100 mg x kg(-1) to mice respectively. The sedative effects were determined by autonomic action test. The analgesic effects were observed by the twisting test induced by acetic acid. The antidepressant effects were investigated by the forced swimming test. The anti-inflammatory effects were studied by auricle swelling induced by xylene.

RESULTEtOAc extracts could significantly lower autonomic action times, reduce squirming induced by acetic acid accumulated immobility time in forced swimming and auricle swelling values by xylene. But EtOH extracts could only decrease autonomic action times and accumulated immobility time.

CONCLUSIONThe active ingredients with sedation, analgesia, antidepressant and anti-inflammation in P. boehmeriaefolium var. tonkinense exist mainly in the EtOAc extracts.

Acetates ; Analgesics ; pharmacology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Antidepressive Agents ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hypnotics and Sedatives ; pharmacology ; Male ; Mice ; Piper ; chemistry ; Plants, Medicinal ; chemistry

Twelve compounds were isolated from the aerial parts of Achillea alpina by column chromatographies on silica gel, Sephadex LH-20, and semi-preparative HPLC. The structures were elucidated on the basis of spectral analysis. The compounds were identified as pellitorine(1), 8,9-dehydropellitorine(2), (E,E)-2,4-undecadien-8, 10-diynoic acid isobutylamide(3), (E,E)-2,4-tetradecadien-8,10-diynoic acid isobutylamide(4),sintenin(5), 4',5,7,8-tetramethoxyflavone(6), chrysoplenetin(7), formononetin(8), aurantiamide(9), asperglaucide(10), artemetin(11), and eupatorin(12). compounds 1-5 were isolated from this plant for the first time, and compounds 6-10 were isolated from the genus Achillea for the first time.
Achillea ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo compare therapeutic effects, safety and tolerance of TCM, western medicine and integrated Chinese and western medicine for treatment of acute lumbosacral pain induced by prolapse of lumbar intervertebral disc.

METHODSNinety cases were randomly divided into 3 groups, 30 cases in each group. They were treated respectively with western medicine, TCM and combined TCM and western medicine, and the pain intensity, activity, muscular tension, and other indexes were monitored after 7 days and 30 days of treatment.

RESULTSAfter treatment of 7 days, the combined treatment group in improvement of VAS scores of lumbosacral pain and radiating pain of the lower limbs was superior to the TCM group with no significant difference between the two groups, and in improvement of VAS scores of lumbosacral pain and radiating pain of the lower limbs, Lasegue's sign, activity of spinal column (Schober test and distance from finger tip to floor), etc. were superior to the western medicine group (P < 0.05). After treatment of 30 days, there was no significant differences in the therapeutic effect among the 3 groups. The patients in the 3 groups had good tolerance with no severe adverse reaction.

CONCLUSIONCombined TCM and western medicine treatment has rapid effects and definite therapeutic effect in alleviating pain, improving activity for acute lumbosacral pain induced by prolapse of lumbar intervertebral disc.

Benzodiazepines ; therapeutic use ; Humans ; Intervertebral Disc Displacement ; therapy ; Ketoprofen ; therapeutic use ; Lumbar Vertebrae ; Medicine, East Asian Traditional

Background The surgery for femtosecond laser created laser in situ keratomileusis(LASIK)flaps has made great progression recent year,but the postoperative corneal wound healing and regeneration of nerve fibers after surgery are closely concerned.Objective This study was to compare and analyze the clinical outcomes between FEMTO LDV femtosecond laser flap and mechanical microkeratome Hansatome flap in LASIK.Methods A prospective case-controlled study was designed.The serial 38 myopic eyes of 38 patients were included from March through July,2010 in Henan Armed Police Force General Hospital.The patients were randomized into FEMTO LDV femtosecond laser assisted flap group(20 patients/20 eyes)and mechanical microkeratome Hansatome assisted flap group(18 patients/18 eyes)with the matched age,gender and refraction of spherical equivalent.HRT Ⅲ examinations were performed before surgery,1 week,1 month and 3 months after surgery to compare the morphological changes atthe center and margin of the flaps,and evaluate the similarities and differences of cellular morphology after surgery between the two approaches.Written informed consent was obtained from each patient prior to this medical trial.Results The best corrected visual acuity was ≥ 1.0 and the refract diopter was similar in both groups(+0.21 D±0.48 D and-0.04 D±0.54 D)1 month after LASIK.The corneal thickness was insignificant increased in the first week after LASIK,and the density of shallow stromal cells was decreased in 1 week,1 month and 3 months compared with pre-operation in the femtosecond laser assisted flap group(t =-27.99,-25.49,-28.87,P ＜ 0.01).In the Hansatome assisted flap group,significantly thickened corneal epithelium was seen in the first week after LASIK compared before LASIK(56.73 μm±2.47 μm versus 51.16 μm±1.11 μm)(t=9.29,P＜0.05),and the density of shallow stromal cells was decreased in 1 week,1 month and 3 months compared with pre-operation in the Hansatome assisted flap group(t =-17.57,-14.13,-19.63,P =0.00).The density of high reflective interface particles in cornea was lower in 1 week,1 month and 3 months after LASIK in the femtosecond laser assisted flap group than that in the Hansatome assisted flap group,showing significant differences between them(t =-13.505,-11.900,-14.084,P＜0.01).The active stromal cells were seen beneath the interface in both groups in the first week and gradually decreased after that time.Intact corneal nerve fibers were found in the femtosecond laser assisted flap group,but those in the Hansatome assisted flap group were shorter and smaller 3 months after LASIK.At 3 months after surgery,the flap margin showed stromal higher reflection and irregular secondary fibrosis in the femtosecond laser assisted flap group,and in contrast,the flap margin had the appearance of a unclearly identified fibrotic scar in the Hansatome assisted flap group.Conclusions Compared with the LASIK and Hansatome assisted flap,the LASIK with FEMTO LDV flap shows earlier nerve fiber regeneration and greater fibrotic scarring,which imply a good wound healing process in the LASIK with FEMTO LDV flap.

Objective To explore the influence of coal-arsenic exposure on human T cells proliferation and its mechanism.Methods Blood samples colleoted from individuals which lived in arsenism area of coal-burning type and non-arsenism area in Guizhou Province were divided into exposed group(17),mild(35),moderate(38) and severe arsenism group(19)and control group(35)according to Diagnosis Smndard for Endemic Arsenism (WS/T 211-2001).T cell stimulation index wag determined by methyl thiazolyl tetrazolium(MTT)colorimetric method.The intracellular Ca2+ exponential(IECa2+)in peripheral blood mononuclear cell(PBMC)was analyzed by Fho-3/AM dye and flow cytometry.DNA binding activity of actively T cells nuclear factor(NF-AT)in PBMC was evaluated by electrophoretie mobility shift assay(EMSA).Results Concanavalin A(ConA)stimulation decreased the T cells stimulation indexes in exposed group,mild,moderate and severe arsenism groups(1.315±0.962, 1.611±1.224,1.114±0.545,1.289±0.875)compared with control group(2.322±1.241),all the differences being statistically significant(P＜0.01).After stimulated by anti-CD3 monoclonal antibody(McAb),the T cells stimulation index in exposed group,mild,moderate and severe arsenism group(0.997±0.177,1.103±0.291,1.007±0.221, 0.957±0.205) were lower than that of control group(1.842±0.429,P ＜ 0.01 ). IECa2+ of PBMC after treated by anti-CD3 McAb in mild,moderate and severe arsenism group( 110.130±49.637,92.429±31.191,77.640± 35.372) were lower compared with control group(145.986±59.450,P ＜0.01 ). Moreover,IECa2+ in moderat and severe arsenism group were lower than exposed group(121.337±46.410,P ＜ 0.05). DNA binding activity of PBMC NF-AT in mild,moderate and severe arsenism group(1.354±0.446,1.290±0.291,1.159±0.411 ) were lowered than that of control group(1.722±0.291,P ＜ 0.01) and exposed group(1.611±0.294,P ＜ 0.05). Conclusions The coal-arsenic exposure can reduce the human T cells stimulation indexes,IECa2+ in PBMC and the DNA binding activity of NF-AT. It suggest that arsenic may suppress the proliferation ability of human T cells,which may be partly related to the influence of arsenic on T cell receptor(TCR)/CD3 signal transduetion pathway.

Objective To investigate the activation of T lymphocytes in human peripheral blood and the signaling molecules in protein kinase C/nuclear factor KB(PKC/NF-κB) pathway expressivity or activity changes in human peripheral blood mononuclear cells(PBMCs) exposed to coal-arsenic,to explore the role of PKC/NF-κB signal pathway in activation of T cells in human exposed to coal-arsenic. Methods Blood samples were collected from individuals who lived in arsenism area of coal-burning in Guizhou province, and were divided into asymptomatically exposed group (12),mild arsenism group (33),moderate arsenism group (34) and severe arscnism group (15) according to Diagnosis Standard for Endemic Arsenism (WS/T 211-2001). The individuals who lived in non-arsenism area were control group(27). The ratio of activated T ceils was analyzed by flow cytometry. DNA binding activity of NF-κB in PBMCs was evaluated by electrophoretic mobility shift assay(EMSA). The expression of PKCθ and phospho-PKCθ(pPKCθ) in PBMCs were detected with western blotting analysis. Results The ratio of activating T cells in asymptomatically exposed group[(21.76±15.31)%],mild arsenism group[(18.41±11.36)%],moderate arsenism group[(17.78±11.93)%]and severe arsenism group[(18.79±13.38)%]were all higher than that of control group[(3.19±2.12)%],the difference among all groups being statistically significant(F = 7.893,P < 0.05). DNA binding activity of PBMCs NF-κB in asymptomatically exposed group,mild arsenism group,moderate arsenism group and severe arsenism group(1.49±0.24,1.58±0.30,1.57±0.34,1.51±0.16) were higher than that of the control group(1.30±0.17),the difference being statistically sign/ficant(P < 0.05 or < 0.01). The expression of PBMCs pPKCθ in mild arsenism group,moderate arsenism group and severe arsenism group(0.64± 0.14,0.64±0.27,0.62±0.12) were all lower than that of the control group(0.93±0.20),the difference being statistically significant(P < 0.05). There were significant negative correlations between the expression of pPKCθ and the activity of NF-κB(r =-0.565,P < 0.01). There were significant positive correlations between the activity of NF-κB and the ratio of activating T cells(r = 0.546,P < 0.01). Conclusion Coal-arsenic enhances the DNA binding activity of NF-κB,reduces the expression of PBMCs pPKCθ in human PBMCs and up-regulates the activity of T cells. It suggests that the PKC/NF-κB signal might be one of transduction pathway via activating of T cells by coal-arsenic.

Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P ＜ 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P ＜ 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P ＜ 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P ＜ 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P ＜ 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P ＜ 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.

Objective To determine the protein expression of P14ARF,MDM2 and mutant type P53 (P53mt) in skin specimens of coal-burning-type of endemic arseniasis patients and to reveal the molecular mechanism of the disease.Methods Sixty skin specimens from 60 endemic arseniasis patients including 35 of skin lesions patients,19 of precancerous lesion and 6 of skin cancer and 9 normal skin specimens from non-cancer patients were studied.Expression of P14~,MDM2 and P53mt was evaluated by immunohistochemistry using corresponding monoclonal antibodies.Results There was significant difference in the positive rates of P14ARF,MDM2 and P53mt among the 4 groups(x2 =9.39,6.21,20.64,all P ＜ 0.05).The positive rates of P14ARF in precancerous lesion and skin cancer specimens were 46.1％ (6/19) and 33.3％ (2/6),respectively,which were significantly lower than that of the normal skin specimens [88.9％(8/9),all P ＜ 0.05].Decreased expression of P14ARF was correlated with the development of dermopathy (P ＜ 0.05).The positive rates of MDM2 and P53mt in skin lesions,precancerous lesion and skin cancer specimens were 54.2％ ( 19/35 ),63.2％ (10/19),66.7％ (4/6) and 25.7％(9/35),73.7％(14/19),83.3％(5/6),respectively,which were significantly higher than those of the control (0,0,all P＜ 0.05).The expression of MDM2 and P53mt increased with the development of dermopathy(all P ＜ 0.05).Conclusions P53mt protein in skin tissue of coal-burning-type of endemic arseniasis patients is over expressed.Abnormal expression of P14ARF and MDM2 may be one of the reasons lead to abnormal cell cycle control disorders and may play a role in the development of endemic arseniasis.