0.05). Flow cytometry analysis by diff erent gates showed the transfection ratio was high in BMSCs in the period of pro d uctive metabolism. The mRNA expression of CAR in P3, P6 and P8 was similar, and the same change was observed in the protein expression of CAR in P3 and P8 BMSCs . CONCLUSION: Ad-CMV-GFP is transferred to BMSC effectively and sustained about 28 days. It is suspected that BMSCs in mitotic phase are easy to be transferred by Ad-CMV-GFP and different passages of BMSCs from P3 to P8 BMSCs can be as high-effectively g ene vehicle.

Objective To confirm an alternative method for primary culture of osteoblasts through comparison of two primary culture methods. Methods Calvarias were dissected from newborn Kunming mice, osteoblasts were isolated with serum-containing collagenase digestion method and sequential digestion method respectively. The osteoblasts were observed under invert microscope, the growth curve was made with the application of MTT method, the rate of living osteoblasts was counted with trypan blue method respectively. Results Compared with the serum-containing collagenase digestion method, the sequential digestion method presented higher production of osteoblasts, higher cell survival rate (P

Objective To explore the effects of different concentrations of fluoride, aluminum alone and in combination exposure on mice parietal bone cell subclone 14 (MC3T3-E subclone 14), and to elucidate the pathogenesis of endemic fluorosis. Methods The proliferation of MC3T3-E1 cells exposed to 10-9 -10-3 mol/L NaF alone, 50 ?mol/L NaF and 5 ?mol/L AlCl3 alone and in combination ,was measured by CCK-8, and the change of cell cycle was measured by flow cytometry after treatment with various concentrations of fluoride and aluminum. Results Fluoride alone did not promote osteoblast MC3T3-E1cells proliferation, higher concentration fluoride inhibited MC3T3-E1 cells proliferation. Fluoride and aluminum combined exposure (50 ?mol/L NaF +5 ?mol/L AlCl3) stimulated proliferation of MC3T3-E1 cells (P

Objective To observe and analysis the features of images of fundus fluorescein angriography (FFA) in low-perfused retinopathy caused by cephalo-cervical peripheral vascular stenosis or occlusion. Methods The results of FFA of 27 patients diagnosed with carotid artery stenosis or occlusion by digital subtraction angiography (DSA) and examination of Doppler and vascular-pulsation were retrospectively analyzed. Result All of the patients had a delayed arm-retinal circulation duration from 20.0 to 81.08 seconds with the mean of 32.1 seconds; a delayed retinal arteriovenous filling duration from 6 to 64.0 seconds with the mean of 24.2 seconds. Delayed arm-retinal circulation duration and retinal arteriovenous filling duration in 10 cases (37.0%); microangioma, vascular wall staining, nonperfused capillary area in 11 (40.7%); and anterior ischemic syndrome in 6 (22.2%) were found. In the 6 patients with anterior ischemic syndrome, 4 cases had narrow retinal artery, segmental changes of blood stream, vascular atresia, and abnormal arterio-venous anastomosis, and 2 cases had bold vascular loops. Conclusions The main manifestations of FFA in patients with low-perfused retinopathy are malperfusion and retinal ischemia, whose degrees relate to the extend of carotid artery stenosis or atresia, and the process of the disease.Serious retinal ischemia may combined with anterior ischemic syndrome.

OBJECTIVE:To establish the Microbial limit test(MLT) for Danggui funing drop pills.METHODS:The recovery rates of 5 tested strains treated by Danggui funing drop pills were detected and the MLT method for the control bacteria was validated.RESULTS: Danggui funing drop pills exhibited strong inhibitory effect on staphylococcus aureus and bacillus subtilis,but showed no effect on escherichia coli,candida albicans and aspergillus niger.CONCLUSION: By membrane-filter procedure,the antibacterial effect of Danggui funing drop pills can be eliminated and the bacterial count can be conducted;however,by routine method,the test of control bacteria is feasible.

Objective To study the effects of fluoride on longitudinal growth and pathological changes of cultured rat metatarsal bone rudiments.Methods Twenty-four neonatal SD rats were divided into four groups according to the random number table,then the second,third and fourth metatarsal bone rudiments were surgically isolated.The left metatarsal bone rudiments were cultured in α-MEM without F-as control group and the right metatarsal bone rudiments from the same rat were cultured in α-MEM with 1 × 10-7,1 × 10-6,1 × 10-5 and 1 × 10-4 mol/L F-.The length and width of the mineralized area of metatarsal were measured on day 0,day 1,day 4 and day 7,respectively,and the pathological changes of metatarsal bone rudiments were observed by the routinely paraffin-embedded sections method on day 7.Results On day 7,the length of the mineralized area was significantly lower of right metatarsal bone [(240.5 ± 139.3)μm] than the left metatarsal bone [(394.1 ± 173.9)μm,t =4.37,P ＜ 0.01] in the 1 × 10-4 mol/L F-group,but the width of the mineralized area [(239.9 ± 119.4)μm] was not different significantly compared to the left metatarsal bone [(223.3 ± 99.9)μm,t =0.44,P ＞ 0.05].The relative vertical growth rate of the mineralized area on day 4 was significantly lower of right metatarsal bone [(2.43 ± 1.44)％] than left metatarsal bone [(8.34 ± 1.74)％,t =3.21,P ＜ 0.01] in 1 × 10-4 mol/L F-group,and the difference was also significant on day 7 [(16.16 ± 2.87)％ vs.(26.52 ± 4.46)％,t =3.13,P ＜ 0.01].Toluidine blue staining results showed that the thickness of cartilage cells of proliferation and hypertrophic layers was significantly lower of right metatarsal bone [(111.33 ± 27.29),(125.33 ± 30.08)μm] than left metatarsal bone [(127.33 ± 38.36),(160.50 ± 42.73)μm,t =4.82,5.81,all P ＜ 0.01] in 1 × 104 mol/L F-group.The ratio of proliferative and hypertrophic layers was significantly higher of right metatarsal bone (0.93 ± 0.36) than left metatarsal bone (0.83 ± 0.32,t =4.42,P ＜ 0.01) in 1 × 10-4 mol/L F-group.Conclusions Our findings indicate that excessive fluoride could cause longitudinal bone growth inhibition.Such growth inhibition is mediated by decreased chondrocyte proliferation and differentiation and the disproportion of proliferation and differentiation.

Objective To explore the clinical feature and therapeutic strategy for children with thrombotic microangiopathy (TMA). Methods The clinical manifestation, auxiliary examination and treatment of 24 cases of children with TMA was analyzed retrospectively. Hemolysis urine toxin syndrome (HUS)occurred in 22 cases, and thrombotic thrombocytopenic purpura (TTP) occurred 2 cases. Results Sixteen cases was onset from May to July,and 8 cases was onset from September to November. Typical HUS (D+HUS) was 8 cases, and atypical HUS (D-HUS) was 14 cases. In 22 HUS children, 18 cases were given hemodialysis or peritoneal dialysis treatment. The illness were significantly improved, and the platelet count and renal function fully recovered normal. But 1 case appeared neurological symptoms such as headache, facial paralysis on one side, gastrointestinal bleeding, and fever, after getting better. Eventually the patient died of disseminated intravascular coagulation. Two cases of TTP were given plasma exchange , anti-infection, large dose of methylprednisolone and anti- platelet aggregation treatment . After treatment, the level of hemoglobin and blood platelets returned normal and consciousness was recuperated. Conclusions HUS and TTP are similar in pathogenesis and clinical manifestation,and it is necessary to be differentiated. Early diagnosis and proper treatment is the key to save the life of children with TMA. As soon as the diagnosis is clear, hemodialysis or peritoneal dialysis treatment should be given to HUS, and plasma exchange to TTP,to quickly control the condition and improve the clinical symptoms.

Objective To investigate the clinic application of micro-implant anchorage in the treatment of maxil-lary protrusion malocclusion. Methods Twenty-two patients,aged 18 to 25 years old,with maxillary protrusion were divid-ed into two groups:experimental group and control group with 11 patients in each group. All patients were treated with ex-traction. Micro-screw palatal implant was used in the cases of experimental group as orthodontic anchorage ,and traditional anchorage composed of extraoral arch used in the cases of control. The cephalometric films were measured before and after treatment. Statistical methods were utilized to analyze the morphological changes of facial profile and hard tissues in both groups. Results The values of U1-NA(mm:3.08±1.18 vs 8.15±3.05) and U1-SN(101.90°±3.50° vs 117.90°±6.05°) were sig-nificantly decreased after treatment compared with those before treatment in the experimental group ( P<0.01). The value of U1-L1(123.98°±5.78°vs 103.89°±8.95°) was significantly increased after treatment (P<0.01). In control group, the values of U1-NA (mm:5.01±1.34 vs 9.12±2.13) and U1-SN(101.90°±3.97° vs 114.87°±7.69°)were significantly decreased after treat-ment. The values of U1-L1(126.01°±3.12°vs 112.98°±5.98°) and U6-PtPNS(mm:21.45±2.43 vs 18.36±2.19)were significant-ly increased after treatment (P<0.05). The value of U1-L1(19.48°±8.90° vs 13.01°±5.90°) was significantly changed in exper-imental group than that of control group, but the value of U6-PtPNS(mm:0.90±0.29 vs 3.78±0.12)was significantly changed in control than that of experimental group (P<0.01). Conclusion The maxillary protrusion malocclusion with micro-im-plant anchorage can be used as treatment for patients with maxillary protrusion that needs strong anchorage.

BACKGROUND:Studies addressing reconstruction of tooth tissue engineering have shown that tooth structure can be constructed using tissue engineering technology. Tooth root and its periodontal attachment are critical for tooth survival and functions, based on which, whether we can target root tissues with simple structure for tissue engineering construction by bypassing a complex dental tissue engineering concept with the structural integrity?
OBJECTIVE:To construct a tissue-engineered dental root by seeding dental papil a cells, as seed cells, into poly(lactic-co-glycolic acid)/sodium alginate hydrogel.
METHODS:Rabbit dental papil a cells were isolated and cultured. The cells were then mixed with 1%sodium alginate hydrogel at a final density of 6×109/L. The cellsuspension was seeded into a poly(lactic-co-glycolic acid) scaffold with predetermined shape of human tooth and solidified with calcium chloride. Final y, the cel-scaffold composites were subcutaneously implanted into the back of nude mice. The specimens were harvested after 4 and 8 weeks respectively and processed for gross inspection, X-ray and CT examination and histological observation.
RESULTS AND CONCLUSION:The newly formed tissue kept the original shape of human dental root 4 and 8 weeks post-implantation. After 4 weeks of implantation, the specimen density was low;the root implants appeared to be incompletely mineralized, alginate hydrogels were degraded, but the copolymer scaffold was not degraded;a number of dentin-like structure appeared, and a fibrous membrane structure was visible on the surface of specimens paral el to the root surface, but the structure was not continuous, and no pulp cavity formed. After 8 weeks, the newly formed tissue was highly mineralized close to root tissue of the nature tooth;the copolymer scaffold was mostly degraded;specimens appeared to have a large number of mature dentin-like structure, and form continuous fibers membrane on the surface paral el to the root surface, below which, cementum-like structure formed. Artificial dental root with biological y similar structures of human dental roots can be constructed using the method of tissue engineering.

Objective To study the value of the semiquantitative-parameter analysis of wash out index of time-intensity curve (Swash-out) in evaluating the therapeutic effect of neoadjuvant chemotherapy for locally advanced breast cancer (LABC).Methods Fifty-nine women with LABC underwent dynamic contrast enhancedt MRI examination before chemotherapy,after the 2nd cycle and the 4th cycle of chemotherapy.All patients were divided into major histological response group (MHR) and non-major histological response group (NMHR) according to the final pathologic response.Swash-out and the variancetrends of Swash-out before NAC,after the 2nd cycle of NAC and after the 4th cycle of NAC were compared in each group and between the two groups.According to the gold standard of Miller & Payne criterion,Receiver operating characteristic curve (ROC) analysis was performed to evaluate the predicting effect of Swash-out for NAC response,and to compare it with Semi-quantitative TIC curve indicators Smax (steepest slope) and PPE (peak percent enhancement).Results Fifty-nine patients of LABC patients were divided into a MHR group of 34 patients and a NMHR group of 25 patients.Swash before NAC of MHR group was-16.99 (-56.72-41.20),Swash-out after the 2nd cycle of NAC was 5.66(-69.45-53.08),Swash-out after 4th cycle of NAC was 15.95 (-7.80-54.23).Swash-out before NAC of NMHR group was-23.08 (-64.24-34.39),Swash-out after the 2nd cycle of NAC of NMHR group was-23.01 (-52.72-28.70),Swash-out after 4th cycle of NAC of NMHR group was-11.45 (-50.49-50.93).Swash-out variance rate of MHR group after the 2nd and the 4th cycle of NAC were-1.18 (-31.32-60.86) and 1.50 (-86.27-3.61),respectively.Swash-out variance rate of NMHR group after the 2nd and the 4th cycle of NAC were-0.28(-3.24-9.46) and 0.27 (-5.34-3.11),respectively.Swash-out was not significantly different between the two groups before NAC (Z =-0.97,P ＞0.05).Swash-out and Swash-out variance rate of MHR group after the 2nd cycle of NAC were significant higher than that of NMHR group (Z =-3.97 and-3.02,P ＜0.01).Swash-out and Swash-out variance rate of MHR group after the 4th cycle of NAC were significant higher than that of NMHR group (Z =-3.96 and-3.16,P ＜ 0.01).Area under curve (Az) after the 2nd and the 4th cycle of NAC were 0.805 and 0.804,respectively,and no significant difference was found between them (Z =0.019,P ＞0.05).Diagnostic cut-off points were-8.670 for the 2nd cycle of NAC and 4.105 for the 4th cycle of NAC.Diagnostic sensitivity was 79.42％,specificity was 76.00％ and Youden index was 0.554,for after the 2nd and the 4th cycle of NAC.Conclusion Swash-out of TIC curve before NAC cannot predict the response of NAC,Swash-out of TIC curve after the 2nd cycle of NAC and after the 4th cycle of NAC are efficient in predicting the response of NAC.