Objective To investigate the effect of triiodothyronine (T3) on neurogenesis in after bilateral common carotid artery occlusion in adult rats. Methods Fifteen adult male Sprague-Dawley (SD) rats were randomly divided into sham operation, 2-vessel occlusion (2VO) and T3 intervention groups (n = 5 in each group ). A chronic cerebral ischemia model was established by the bilateral common carotid artery permanent ligation. Seven days after modeling 5-bromo-2'-deoxyuridine (BrdU) was injectied intraperitoneally and the new proliferating cells were labeled. The BrdU positive cells in SVZ were observed using immunofluorescence staining.Results The numbers of BrdU positive cells in the sham operation, 2VO, and T3 intervention groups were 174. 8 ± 18. 5, 325.0 ± 51.0, and 499. 4 ± 30. 8, respectively. There were significant differences among all the groups (F = 101.68, P ＜ 0. 001 ). The numbers of BrdU positive cells in the T3 intervention group were significantly higher than those in the 2VO and sham operation groups (all P ＜0. 001 ). The numbers of BrdU positive cells in the 2VO group were also significantly higher than those in the sham operation group (p ＜0. 001). Conclusions Thyroid hormones may promote neural progenitor cell proliferation in SVZ in adult 2VO rats.

Objective　To investigate the different expression of actin, myosin Ⅱ in hypertrophic scars, keloids and normal skins, and to understand the relationship of actin, myosin Ⅱ and the scar contracture. Methods Fifteen cases with hypertrophic scars, 10 cases with keloids and 15 cases with normal skins were chosen randomly. The expression of actin and myosin Ⅱ were detected by immunohistochemical method in the hypertrophic scars, keloids and normal skins. The fibroblasts isolated from three types of tissue were cultured in vitro, then actin and myosin Ⅱ in three different fibroblasts were measured using flow cytometry. Results　The immunohistochemical staining of myosin Ⅱ in hypertrophic scars was positive, while the staining in keloids and normal skins were negative. The positive rate of myosin Ⅱ expression in hypertrophic scars, keloids and normal skins were (95.11±2.78)%， (16.86±7.11)%, and (5.31±1.79)% respectively. There were significant difference between keloids and the two others(P＜0.01). The actin expression in three difference tissues were positive, there were no significant difference in hypertrophic scars, keloids and normal skins(P>0.05). The positive rate of actin expression in hypertrophic scars, keoids and normal skins were(77.77±15.43)%, (88.89±10.29)%, and (82.92±13.48)% respectively, and there were no significant difference(P>0.05). Conclusion　Myosin Ⅱ may play an important role in the scar contracture. Actin is the contractile protein of cell, it plays important role in cellular movement. Actin is necessary protein in the cell.

Objective To investigate the effect of electroacupuncture (EA) on oocyte quality in patients with polycystic ovary syndrome (PCOS) undergoing in-vitro-fertilization and embryo transfer (IVF-ET).Method Totally 217 patients with PCOS undergoing IVF-ET were randomized into two groups by the random number table, 119 patients in an EA group and 98 in a control group. In addition to the long-term ovarian hyperstimulation given to the two groups, electroacupuncture was involved in the intervention in the EA group. The spindle, quality of eggs, and pregnancy result were observed and compared.Result The high-quality embryo rate was significantly higher in the EA group than the control group (P<0.05), and compared to the control group, the pregnancy rate was 8.36% higher in the EA group; it’s found that the number of eggs with spindle located between 11 and 1 o’clock was in a significant positive correlation with the level of E2 on the HCG administration day and the high-quality embryo rate (P<0.01); the EA group had a markedly higher ratio of eggs with spindle located between 11 and 1 o’clock compared to the control group (P<0.05); electroacupuncture markedly reduced the dosage and duration of using Gn in the PCOS patients (P<0.05). Conclusion It’s proven the relation between spindle and the quality of eggs; electroacupuncture can enhance the quality of eggs and the pregnancy rate in the PCOS patients.

Agriculture ; Humans ; Occupational Exposure ; adverse effects ; analysis ; Radiation Dosage ; Radon ; adverse effects ; analysis ; Seasons

Objective To investigate the expression of CD26 (dipeptidyl peptidase 4) in the kidney tissues of diabetic rats and the effects of mycophenolate mofetil (MMF) on the renal CD26 expression.Methods Wistar rats were randomly divided into three groups:normal control group (NC group,n=7),diabetic model group (DM group,n=7) and MMF-treated group (MMF group,n=7).Wistar rats were fed with high-sucrose-high-fat diet and injected with streptozotocin into abdominal cavity to induce diabetes.Sixteen weeks later,blood glucose (BG),blood urea nitrogen (BUN),serum creatinine (Scr),renal hypertrophy index (kidney weight]body weight) and 24 hour urinary protein (24Upro) were measured.The number of CD37CD4+ T cells in renal tissues were measured through flow cytometry.The expression of CD26 in kidney was examined by using Western blotting and immunohistochemistry.Results Compared with NC group,BG,BUN,Scr,kidney weight/body weight,24Upro were significantly increased in DM group (P ＜ 0.05).Except BG and kidney weight] body weight,the above-mentioned parameters were lower in MMF group compared with that in DM group (P ＜ 0.05).Intrarenal CD37CD4+ T cells were significantly up-regulated in DM group compared with that in NC group (P ＜ 0.01).CD26 in renal tissue was mainly expressed in T lymphocytes of renal interstitium.CD26 expression in DM group was significantly higher than that in NC group,and also higher than that in MMF group (P ＜ 0.05).In DM group,CD26+ T lymphocytes infiltration of renal interstitium was positively correlated with 24Upro (r2=0.770,P ＜ 0.05).Conclusions CD26 is related with diabetic nephropathy.MMF maybe inhibit T lymphocytes infiltration to reduce the expression of CD26 in renal interstitium,thus protecting the kidney function.

OBJECTIVETo observe the effect of electro-acupuncture (EA) treatment on the oocyte quality in polycystic ovarian syndrome (PCOS) patients undergoing in vitro fertilization-embryo transfer (IVF-ET).

METHODSTotally 217 PCOS patients undergoing IVF-ET were assigned to two groups by random digit table, the EA group (119 cases) and the control group (98 cases). All patients received long program ovarian hyperstimulation with gonadotropin-releasing hormone agonist. Patients in the EA group received EA treatment in the process of controlled ovarian hyperstimulation till the oocyte retrieval day. The position relation of the spindle to the polocyte, the number of retrieved oocytes, the fertilization rate,the cleavage rate,the high quality embryo rate, the ovarian hyperstimulation syndrome (OHSS) incidence rate, the clinical pregnancy rate, the early abortion rate, the gonadotropins (Gn) dose and time, levels of estradiol (E2), progesterone (P), and luteinizing hormone (LH) on the day of human chorionic gonadotropin (HCG) were observed between the two groups.

RESULTSThe ratio of oocytes in which the meiotic spindle deviation angle was < 60 degrees to the all oocytes was obviously higher in the EA group than in the control group (P < 0.05). The oocytes in which the meiotic spindle deviation angle was < 60 degrees was positively related to level of E2 on the HCG day and the high quality embryo rate (r = 0.19,P < 0.01). Compared with the control group, the high quality embryo rate increased significantly (P < 0.05), the dose and days of Gn decreased significantly (P < 0.05) in the EA group. The clinical pregnancy rate was improved by 8.36%.

CONCLUSIONSThe spindle was positively correlated with the oocyte quality. EA could improve the quality of oocytes and the clinical pregnancy rate in PCOS patients undergoing IVF-ET.

Acupuncture Therapy ; methods ; Embryo Transfer ; Estradiol ; Female ; Fertilization in Vitro ; Gonadotropin-Releasing Hormone ; Gonadotropins ; Humans ; Luteinizing Hormone ; Oocytes ; Ovarian Hyperstimulation Syndrome ; Polycystic Ovary Syndrome ; therapy ; Pregnancy ; Pregnancy Rate ; Progesterone

BACKGROUND:Alveolar bone remodeling and sustained absorption due to tooth extraction seriously affect the implanting conditions and morphology of hard and soft tissue in implant zone. OBJECTIVE:To evaluate the effect of nano-hydroxyapatite/polycaprolactone electrospinning scaffolds to improve the osteogenic effect of bone defects around immediate implants. METHODS:Tissue-engineered bone was prepared by combining canine bone marrow mesenchymal stem cels with nano-hydroxyapatite/polycaprolactone electrospinning scaffold. Bilateral mandibular second premolars from six dogs were extracted mandibular second premolar, and an immediate implant was placed in the mesial fossa of the mandibular second premolar. Three-wal bone defects was made buccaly using titanium nails, then tissue-engineered bone and Bio-Oss bone powders were implanted bilateraly covered by colagen membranes (Bio-Gide). Imageology examination was performed to measure bone gray levels immediately, 4, 8, 12 weeks after surgery. After 12 weeks, the mandible was removed completely, toluidine blue staining was used for observation of microstructure, new bone formation, bone morphology and implant osseointegration. RESULTS AND CONCLUSION: Between the two groups, there was no difference in bone mineral density at each time point after surgery, indicating that the effects of the two materials to promote bone regeneration process are basicaly the same. After implantation, the dense lamelar bone formed in the bone defect region of tissue-engineered bone group, mature bone cels, Haversian canal, and implant osseointegration were visible. While, in the Bio-Oss group, the lamelar bone was dense, a smal amount of Bio-Oss particles distributed within new bone tissues, fewer bone cels were found, a part of Haversian canal was shown to have blood capilaries, and new bone was in close conjunction with the implant. These findings indicate that the nano-hydroxyapatite/polycaprolactone electrospinning scaffold combined with bone marrow mesenchymal stem cels and Bio-Gide colagen membrane can promote the regeneration of alveolar bone around the implant.

BACKGROUND: Bone morphogenetic protein-2(BMP-2) production of targeted cells is promoted by transfection of adenoviral vectors containing gene, but there are some immune responses. Transfection with plasmid as vector holds promise.OBJECTIVE: To explore the feasibility to construct human bone morphogenetic protein-2 eukaryotic expression vector labeled with green fluorescent protein (GFP).DESIGN: Single sample observation.SETTING: Tianjin Hospital.MATERIALS: The experiment was performed at the Key Laboratory of Hormone and Development, Ministry of Health, Tianjin Medical University from March 2006 to March 2007. pcDNA3.1/CT-hBMP2 plasmid containing full-length hBMP2 gene fragment was provided by Dr. Li; bicistronic eukaryotic expression vector pSELECT-GFPzeo-MCS and Zeo was provided by Invivogen; pTA2(R)-T Easy by Dingguo, China; restriction enzymes BamHI and NheI, T4 DNA ligase by Jingmei Biotech; PCR upstream and downstream primer synthesis and sequencing by Augct, Beijing.METHODS: With pcDNA3.1/CT-hBMP2 as template, hBMP2 target fragment was subcloned by PCR binding with designed specific primers. The fragment was bound with pTA2-T-easy and pSELECT-GFPzeo-MCS, separately, and transfected into DH5 α cells. pSELECT-GFPzeo-hBMP2 containing GFP was obtained after screening.MAIN OUTCOME MEASURES: hBMP2 sequence was identified by PCR; whether hBMP2 was cloned into pTA2-hBMP2 and pSELECT-GFPzeo-MCS was identified by digestion and sequencing.RESULTS: A target fragment of 1 216 bp was obtained by PCR amplification, and cloned into pTA2-T-easy and pSELECT-GFPZeo-MCS. The screening and sequencing results showed that the target fragment was 100% matched with BMP2cDNA sequence (NM-001200) from GenBank.CONCLUSION: hBMP2 eukaryotic expression vector labeled with green fluorescent protein is successfully constructed.

Objective To investigate the effectiveness of femoral troehanter reconstruction and artificial femoral head replacement in treatment of unstable intertrochanterie fractures in the elderly pa- tients. Methods Femoral trochanter reconstruction and artificial femoral head replacement was done on 106 patients with unstable intertrochanterie fractures. There were 45 males and 61 females, at age range of 80-105 years (average 88 years). Most of patients slipt in the room and got fractured. According to the Evans classification, there were 31 patients with type ⅢA fractures, 45 with type ⅢB and 30 with type IV. We used 4 kinds of methods to reconstruct the fracturad imertrochanters : (1) shape of" ∞ " ten- sion band fixation after intertrochanterie fracture reduction;(2) wire loop fixation of the lesser troehanter around proximal femur;(3)defect within the femoral ealear was filled with bone cement and remodeled; (4) for patients with relative intact base of femoral neck, the intertrochanterie fracture was transformed in- to femoral neck fracture and the femoral ealear was fixed with femoral prosthesis stem. Results All the operations continued successfully, with duration of the operation for 45-70 minutes (average 55 minutes). No artificial femoral head dislocation occurred during hospitalization. Of all, 79 patients were followed up for 6-48 months (average 16 months). No Late loosening, dislocation or infections occurred, with total excellence rate of 87.3%. Conclusions For elderly patient with unstable intertrochanterie fractures, reconstruction of femoral intertrochant and artificial femoral head replacement can restore the proximal femoral anatomy, maintain stability of the hip joint and help early functional exeereise, as can reduce ease fatality rate and improve the quality of life.

Objective To investigate the biological behavior and expression of intergrin-linked kinase (ILK) after exposure to transforming growth factor beta 2 (TGF-β32) in human Tenon fibroblasts (HTFs).Methods The primary ceils of HTFs were cultured by tissue attached culture method and identified by immunofluorescence analysis with Vimentin and keratin.The proliferation levels of HTFs induced by different concentrations of TGF-β2 were analyzed by MTT.The α-smooth muscle actin (α-SMA) and ILK mRNA expression were analyzed by quantitative Real-time PCR.The protein expression of α-SMA,ILK and E-cadherin were analyzed by Western Blot,and the protein expression of α-SMA and E-cadherin were also analyzed by immunofluorescence staining.Results MTT analysis showed that the optical density levels of 5.0 μg · L-1 and 10.0 μg · L-1 TGF-β2 were significantly higher than those of 0.1 μg · L 1,1.0 μg · L-1 and the control after exposure for 48 hours and 72 hours,and all these optical density levels were significantly higher than that for 24 hours (all P ＜0.05).The expression of α-SMA and ILK mRNA increased significantly when cells were treated with 5.0 μg · L-1 TGF-β2 for 48 hours in comparison with the control group (all P ＜ 0.05).The protein of α-SMA,ILK and E-cadherin were expressed both in TGF-β2 treated groups and control group,and TGF-β2 up-regulated the expression of them.There were significant differences when compared with the control group (all P ＜ 0.05).Immunofluorescent staining showed that α-SMA and E-cadherin were detected in TGF-β2 treated groups.α-SMA expressed in cytoplasm,while E-cadherin both in cytoplasm and nucleus.Conclusion TGF-β2 can induce the proliferation,transdifferentiation and adhesion of HTFs,and up-regulate the expression of ILK in vitro,suggests that ILK may play a role in the process of scar formation after glaucoma filtration surgery.