BACKGROUND: The proliferation and differentiation of mesenchymal stem cells (MSCs) lack of regulatory functions. Following combining with suitable vectors, MSCs cannot highly effectively proliferate and differentiate, which are keys to prevent MSCs entering the clinic. It is of great importance to effectively regulate the differentiation of stem cells into osteoblasts using pulse electromagnetic field.OBJECTIVE: To investigate the differentiation of mouse MSCs into osteoblasts in vitro following stimulation of pulse electromagnetic field.DESIGN, TIME AND SETTING: The cytological in vitro controlled study was conducted at the Laboratory of Department of Orthopaedics, Puai Hospital of Tongji Medical College, Huazhong University of Science and Technology from May 2004 to October 2007.MATEIRALS: Totally 20 BALB/C mice were supplied by the Experimental Animal Center of Tongji Medical College. Pulse electromagnetic field deviser was designed and made by the Department of Electric Machine, Naval University of Engineering.METHODS: Mouse bilateral femur was sterilely isolated. BMSCs were harvested by the Percoll density gradient centrifugation,and purified and proliferated by the adherent method. Cells at the third passage (2×10~7/L) were incubated in a 6-well plate, and then divided into 4 groups. Cells in the blank control group were incubated in the complete medium. Cells in the pulse electromagnetic field underwent pulse electromagnetic field radiation of 50 Hz, sinusoidal wave, and 1 mT, twice a day, once 30 minutes, with an interval of 12 hours, totally 10 days. Cells in the osteogenic induction group were incubated in the complete medium, supplemented with dexamethasone, sodium glycerophosphate and VitC. Cells in the pulse electromagnetic field + osteogenic induction group were subjected to the same pulse electromagnetic field radiation and then incubated in the complete medium.MAIN OUTCOME MEASURES: The differentiation of BMSCs was measured.RESULTS: Results of alkaline phosphatase staining showed that cells were negative in the blank control group, but weakly positive in the pulse electromagnetic field group, positive in the osteogenic induction group, and strongly positive in the pulse electromagnetic field + osteogenic induction group 10 days following intervention. Compared with the blank control group,absorbance value of type I collagen immunohistochemistry was significantly greater in the osteogenic induction group, pulse electromagnetic field + osteogenic induction group (P < 0.05, P < 0.01).CONCLUSION: Pulsed electromagnetism fields of 50 Hz, waves of sine, with the intensity of 1 mT could promote alkaline phosphatase and type I collagen expression and enhance the differentiation of mouse BMSCs into osteoblasts in vitro.

Child ; Humans ; Practice Guidelines as Topic ; Sepsis ; therapy ; Shock, Septic ; therapy

Objective To investigate the clinical effects of tibiotalocalcaneal arthrodesis (TTCA) using a retrograde intramedullary nail with a valgus curve.Methods At our department,22 patients underwent TTCA using a retrograde intramedullary nail with a valgus curve from June 2009 to January 2014 and were available for complete follow-up.They were 12 men and 10 women,aged from 46 to 79 years (average,62.2 years).There were 3 cases of primary ankle osteoarthritis,9 ones of traumatic arthritis,one of ankle arthritis secondary to severe talar avascular necrosis,3 ones of progressive subtalar arthritis following failed ankle replacement,5 ones of progressive subtalar arthritis following failed ankle arthrodesis,and one of arthritis secondary to equinovarus.The outcome measurements included the American Foot and Ankle Society (AOFAS) ankle-hindfoot scale,EQ-5DTM functional score,radiologic assessment and clinical examination.Results The mean follow-up was 21.3 months (range,from 14 to 38 months).A plantigrade foot and bony union were achieved in all the patients after a mean time of 3.9 months (range,from 2.4 to 6.2 months).Postoperative radiologic results showed a good hindfoot alignment in all the patients.Superficial infection occurred in one patient and loosening of the distal screw in another who asked for removal.The mean postoperative EQ-5DTM functional score and AOFAS ankle-hindfoot score were 69.3 (range,from 20 to 90) and 69.9 (range,from 45 to 85),respectively.Conclusion TTCA using a retrograde curved intramedullary nail may lead to solid fusion and good hindfoot alignment.

OBJECTIVETo observe the effect of magnesium sulfate, Nifedipine Tablet (NT) combined Salvia Injection (SI) on endothelin-1 (ET-1), nitric oxide (NO), thromboxane A2(TXA2), prostacyclin I2(PG2), and hemorheology of preeclampsia patients.

METHODSTotally 704 preeclampsia patients were randomly assigned to the treatment group and the control group, 352 cases in each group. All patients were treated with magnesium sulfate combined NT (on the first day: slow intravenous injection of magnesium sulfate 5 g + intravenous dripping of magnesium sulfate injection 10 g + oral administration of NT 30 mg; on the second and third day, intravenous dripping of magnesium sulfate injection 10 g + oral administration of NT 30 mg), while those in the treatment group were dripped with SI additionally at 20 mL per day for 3 consecutive days. Before and after treatment plasma levels of endothelin-1 (ET-1), nitric oxide (NO), TXA2, PGi2, and hemorheology indicators [such as high blood viscosity (HBV), low blood viscosity (LBV), plasma viscosity (PV), erythrocyte rigidity index (ERI), fibrinogen (FIB)] of two groups were detected.

RESULTSCompared with the same group before treatment, serum levels of ET-1, TXA2, HBV, LBV, PV, ERI, and FIB decreased in the two groups after treatment (P <0. 05), but levels of NO and PG2 increased (P <0. 05). Compared with the control group in the same period, levels of ET-1, TXA2, HBV, LBV, PV, ERI, and FIB decreased in the treatment group after treatment (P <0. 05), but levels of NO and PGI2 increased (P <0. 05).

CONCLUSIONMagnesium sulfate, NT combined SI could effectively regulate the balance of ET-1/NO and TXA2/PGI2, and improve hemorheology of preeclampsia patients.

Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Endothelin-1 ; metabolism ; Epoprostenol ; metabolism ; Female ; Hemorheology ; Humans ; Injections ; Magnesium Sulfate ; administration & dosage ; pharmacology ; therapeutic use ; Nifedipine ; administration & dosage ; pharmacology ; therapeutic use ; Nitric Oxide ; metabolism ; Pre-Eclampsia ; drug therapy ; Pregnancy ; Salvia ; Tablets ; Thromboxane A2 ; metabolism

Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.

Objective To explore the efficacy and safety of adjusting rivaroxaban dose for preventing deep vein thrombosis(DVT) according to extreme body mass in the patients with orthopedic surgery.Methods One hundred and two patients undergoing the first time total hip arthroplasty(THA) from January 2014 to December 2016 included 102 cases of body mass ＜50 kg and 27 cases of body mass ＞100 kg,which were divided into the experimental subgroup and control subgroup respectively.Each subgroup was given different doses of rivaroxaban for 35 d.The coagulation function,hemoglobin,D-dimer change,bleeding events and VTE incidence rate were recorded.Results The extension values of PT and APTT had no statistically significant difference between the experimental group and control group(P＞0.05).Hb and PLT in the ＜50 kg group were decreased,and the incision drainage amount was less than that in the ＜50 kg group,the intra-group comparison in the ＞100 kg group had no statistical difference (P＞0.05).All cases had no bleeding in other sites.The difference of DVT and PE occurrence had no statistical significance(P＞0.05).Conclusion Adjusting the rivaroxaban dose according to the extreme body mass can effectively prevent the DVT occurrence after orthopedic operation and reduces the occurrence rate of adverse drug reactions.

Objective To find out factors related to avascular necrosis (AVN) of femoral head through a retrospective study of patients with femoral neck fractures treated with cannulated screws.Methods Two hundred and thirty cases of femoral neck fracture whn had been treated with cannulated screws in our department from August 1994 to October 2003 were analyzed statistically.Six factors of patient,age,gender,fracture type (Garden clas- sification),reduction method (close or open),the quality of reduction (Garden Index) and time interval from injury to surgery were analyzed statistically with rank sum test and logistic multiple regression.Results Factors that might have an impact on avascular necrosis of femoral head were ranked in the following likelihood sequence: fracture type,quality of reduction,reduction method,time interval from injury to surgery and age.Gender was found to have little impact on the prognosis of femoral neck fracture (P＞0.05).Conclusions Fracture displacement is the most critical factor that influences the prognosis.Anatomical reduction (close or open) must be achieved to re- duce the risk of AVN.Timing of surgery shows its importance.High age will not lead to an increased incidence of AVN.

OBJECTIVETo compare and evaluate the biocompatibility of three kinds of dentin bonding agents Xeno III (XO), Adper Prompt (AP), Single bond2 (SB) through cell culture in vitro.

METHODSThree kinds of dentin bonding agents (XO, AP, SB) were applied on the surface of the dental slices which were 5.0 mm in diameter and 0.5 mm in depth. By immersing the slices into the DMEM culture medium, the maceration extracts were obtained. Normal dental pulps of teenagers were collected and human pulp fibroblast was cultured using tissue explant method. The fifth generation pulp cells were exposed to culture medium containing different concentrations of maceration extracts (100.0%, 50.0%, 25.0%, 12.5%) for 24, 72, 120 h. At last, MTT method was used to evaluate the cytotoxicity of the dentin bonding agents on human pulp fibroblast.

RESULTSThe results showed that all three kinds of dentin bonding systems had cytotoxicity to human pulp fibroblast in different degree in vitro. The cytotoxicity of XO and AP was less than SB. The difference was statistically significant (P<0.05).

CONCLUSIONThe results of cell culture in vitro indicated that total-etching adhesives system has more irritation to pulp than self-etching adhesives system.

Adhesives ; Adolescent ; Dental Pulp ; Dentin ; Dentin-Bonding Agents ; Fibroblasts ; Humans ; Resin Cements

Objective To investigate the influence of blood glucose fluctuation on ser202 phosphorylation sites of tau protein( p-Tau) in hippocampus of diabetic rats; to explore the possible mechanism of blood glucose fluctuation impacting on tau protein hyperphosphorylation. Methods Healthy male Sprague Dawley rats were randomly divided into normal control group ( NC group ) and diabetes group. After diabetic rats model was established, all the diabetic rats were randomly divided into diabetic continuous hyperglycemia group (DC group) and diabetic blood glucose fluctuant group ( DF group). Rats in DF group were given glucose solution intraperitoneal injection twice at regular time everyday. 30 minutes after each intraperitoneal injection, insulin subcutaneously injections were given. Rats in the NC group and DC groups were given the same volume of saline subcutaneous injection. Specimens were collected in 8 weeks, the levels of p-Tau and total tau in rat hippocampus were detected by immunohistochemical staining and Western blotting. The immunoreactive positive products were analyzed by image analysis system. Glycogen synthase kinase-3β(GSK-3β) mRNA was detected by realtime PCR. Results (1) Blood glucose fluctuation of rats in DC and DF group were greater than NC group. And the mean blood glucose, standard deviation of mean blood glucose (SDBG), and large amplitude of glycemic excursion (LAGE) levels were increased significantly compared to NC group, the difference has statistical significance ( all P < 0. 05). Compared with DC group, SDBG and LAGE levels of DF group were higher (both P<0. 05). HbA1C and insulin levels were no difference (P>0. 05). (2) Compared with NC group, the hippocampal p-Tau level of DC group and DF group were increased (P < 0. 05 ); Compared with DC group, the hippocampal p-Tau expression of DF group was increased ( P <0. 05). Compared with DC group, a higher hippocampal GSK-3β mRNA level was found in DF group ( P <0. 05). Conclusions On the basis of diabetes animal model, giving glucose solution intraperitoneal injection and insulin subcutaneously injection 30 minutes later twice at regular time everyday could establish experimental model of diabetic blood glucose fluctuation. Blood glucose fluctuation may aggravate the diabetic rats hippocampal p-Tau. The possible mechanism seems to be an up regulation of the GSK-3β.

Objective To investigate the treatment of subtrochanteric femoral fractures of Sein-sheimer type Ⅲ with prolonged Gamma3 nails and titanium wire fixation in the aged patients. Methods From October 2014 to December 2015, 23 old patients with subtrochanteric femoral fracture were treated with prolonged Gamma3 nails and titanium wire fixation. They were 7 men and 16 women, aged from 62 to 78 years ( 69. 3 years on average ) . By Seinsheimer classification, 14 cases were type ⅢA and 9 type ⅢB. The time from injury to surgery averaged 3. 4 days ( from 2 to 6 days ) . Their operation time, intraoperative bleeding, hospital stay, fracture union time and hip function at the final follow-up were documented. Results Their operation time averaged 1. 3 hours ( from 1. 0 to 1. 6 hours ) , intraoperative bleeding 161 mL ( from 115 to 215 mL ) , and hospital stay 10. 7 days ( from 9 to 12 days ) . They obtained follow-ups from 5 to 12 months ( 9. 3 months on average ) . All the fractures achieved bony union after an average of 15. 4 weeks ( from 12 to 20 weeks ) . The function of the affected hip assessed at the final follow-up according to the Harris scoring was excellent in 15 cases, good in 5 and fair in 3, resulting in an excellent to good rate of 87. 0％. Their follow-ups revealed no deep venous thrombosis of lower limb, fat embolism, coxa vara, refracture, cut-in of the femoral head by the lag screw, or implant breakage. Conclusions In the treatment of senile subtrochanteric femoral fractures of Seinsheimer type Ⅲ, prolonged Gamma3 nails and titanium wire fixation exhibits advan-tages of limited operative trauma, rigid fixation, a high rate of union and limited complications. Additionally, this treatment can increase the accuracy of inserting distal locking nails, shorten operation time and improve functional recovery.