BACKGROUND:Studies have shown that apoptosis is closely related to the pathogenesis of steroid-induced femoral head necrosis. The release of cytochrome c plays a very important role in the process of apoptosis.
OBJECTIVE:To observe the effects of cytochrome c on early steroid-induced femoral head necrosis.
METHODS:Twenty-four healthy adult male New Zealand rabbits at 5 months old were randomly divided into model group and control group (n=12 per group). Models of early steroid-induced femoral head necrosis were established by intragluteal injection of hormone combined with ear vein injection of horse serum. In the control group, rabbits were given ear vein injection of the same amount of physiological saline. At 2, 4 and 6 weeks after model establishment, histopathological changes of bilateral femoral head were observed by optical microscope, and the ratio of empty lacuna was calculated. Apoptosis of osteocytes was determined by TUNEL assay, and apoptotic index was calculated. Immunohistochemical method was used to determine cytochrome c and to calculate cytochrome c-positive expression rate.
RESULTS AND CONCLUSION:(1) The ratio of empty lacuna and apoptotic index:The model of early steroid-induced femoral head necrosis was successful y established in the experiment. Compared with the control group, ratio of empty lacuna, apoptotic index and expression rate of cytochrome c in osteocytes were significantly higher in the model group (P<0.05 or P<0.01). (2) Correlation analysis:Ratio of empty lacuna was significantly positively associated with apoptotic index at various time points in the model group (r=0.856, P<0.01). Expression rate of cytochrome c was significantly positively associated with apoptotic index at various time points in the model group (r=0.824, P<0.01). (3) These findings confirm that cytochrome c-involved apoptosis of osteocytes may play an important role in steroid-induced femoral head necrosis. Expression rate of cytochrome c in osteocytes is remarkably positively associated with the occurrence of steroid-induced femoral head necrosis in rabbits.

Objective To study the anti-asthma mechanism of water extract of Portulaca oleracea L( WEPO) related toβand M receptors.Methods Rabbits of 2.0 -2.5 kg were selected,whose isolated tracheal rings were prepared.The effect of WEPO on tracheal smooth muscle tension was evaluated using tracheal smooth muscle contraction frequency and contraction amplitude as indexes.Aminophylline( Ami) was used as a positive control medicine and propranolol( Pro) as theβreceptor blocking agent to observe the relaxation effect on tracheal smooth muscle of WEPO related to βreceptors. Acetylcholine chloride( AChCl) was used as the agonist for M receptors and atronine( Atr) as the M receptor blocking agent to observe the relaxation effect on tracheal smooth muscle of WEPO related to M receptors.Results ①The contraction frequency and amplitude of isolated tracheal smooth muscle in each dose group of WEPO were significantly lower after treatment.The difference was statistically significant(P <0.05,P <0.01).②The baseline,contraction frequency and amplitude of isolated tracheal smooth in Pro +WEPO group were higher than in WEPO group and the difference was statistically significant (P<0.01).③The baseline, contraction frequency and amplitude of isolated tracheal smooth muscle in AChCl group were significantly higher after treatment(P<0.01)significantly.④The baseline, contraction frequency and amplitude in WEPO +AChCl group were not different from those in AChCl group ( P >0.05 ) .⑤These indexes in Atr+WEPO group were not different those in WEPO group ( P >0.05 ) .Conclusion WEPO has relaxation effect on isolated tracheal smooth muscle of rabbits,which is related toβreceptors rather than M receptors.

Objective To investigate reasonable surgical approaches for lower spine injuries.Methods The study involved 174 patients with lower cervical spine injuries treated with anterior approaches, posterior approaches, or anterior-posterior approaches in our hospital from August 2005 to September 2009. American Spinal Injury Association (ASIA) grading system was used to evaluate the surgical outcome. Results All patients were followed up for average 30 months (6-55 months), which showed that bone union was achieved in 169 patients, with no breakage, loosening or displacement of the internal fixators. There were five deaths. The ASIA grades of 125 patients were improved by 1 or 2 levels ( 1.12levels on average), with an improvement rate of 71.3%. Conclusion Comprehensive and accurate preoperative diagnosis is the basis for choose of correct surgical approaches for lower spine injury. Early and correct surgery is essential for a good prognosis.

Objective To evaluate the effect of IL-18 on the expression of E-cadherin,α-SMA and CX3 CL1 in pancreatic stellate cells (PSCs).Methods The human PSC line HPaSteC was routinely cultured and passaged.Five,25,50 and 100 μg/L IL-18 was used to treat PSCs for 72 h and the untreated PSCs were used as control.Treated and untreated cells were both collected,and RT-PCR and Western blot were used to detect mRNA and protein expression of E-cadherin,α-SMA and CX3CL1 respectively.Results The mRNA expressions of E-cadherin in control group and 5,25,50 and 100 μg/L IL-18 treated group were 1.03 ± 0.17,0.77 ±0.15,0.89 ± 0.12,0.54 ± 0.11 and 0.46 ± 0.06.The mRNA expression of α-SMA were 1.03 ± 0.19,0.85 ± 0.14,1.33 ± 0.22,1.60 ± 0.14 and 1.94 ± 0.09;The mRNA expression of CX3CL1 were 1.01 ±0.08,0.88 ±0.25,0.86 ±0.17,1.58 ±0.26 and 1.83 ±0.13.The mRNA expression of E-cadherin in IL-18 treated group were down-regulated,while the mRNA expression of α-SMA and CX3CL1 were up-regulated,and the differences between control and IL-18 100 μg/L treated group were statistically significant (P ＜ 0.05 or ＜ 0.01).The protein expression of E-cadherin in control group and 5,25,50 and 100 μg/L IL-18 treated group were 1.00 ±0.14,1.14 ±0.04,1.14 ±0.07,0.85 ±0.08 and 0.80 ±0.06.The protein expression of α-SMA were 1.00 ± 0.02,0.77 ± 0.07,1.29 ± 0.02,1.59 ± 0.07 and 1.70 ± 0.02;The protein expression of CX3CL1 were 1.00 ± 0.05,1.03 ± 0.05,1.37 ± 0.06,1.46 ± 0.18 and 1.45 ± 0.12.The protein expression of E-cadherin was down-regulated but no significant differences were observed among different groups.The protein expression of α-SMA was up-regulated and the differences between control group and 25,50 and 100 μg/L IL-18 treated groups were statistically significant (all P ＜0.01).The protein expression of CX3CL1 was up-regulated and the differences between control group and 100 ng/ml IL-18 treated group were statistically significant (P ＜0.05).Conclusions IL-18 can activate PSCs and up-regulate the expression of chemokine CX3CL1.

Objective To study the effect of leech powder on mortality and disability of acute intracerebral hemorrhage at 30 days and 1 year. Methods In a Case-control study,we analyzed the 30-day and 1-year mortality and disability with respect to leech powder treatment status in 59 patients consecutively admitted within one week of intracerebral hemorrhage onset. Confounding factors were compared between treated and nontreated patients. Results About two thirds of the patients received oral leech powder as part of their routine treatment,mean dosage was (2.22?0.42)g/d and mean duration was (10.12?3.39)d. The mortality was 2.4% versus 16.7% (P =0.08) at 30 days and 17.1% versus 44.4% (P=0.03) at 1 year in the leech powder treated and nontreated groups,respectively. Modified Rankin score 5-6 was 43.9% (61.1% in nontreated groups,P =0.45) at 30 days and 17.1% (50% in nontreated groups,P =0.03) at 1 year in treated groups,respectively. Leech powder treatment effect was adjusted for Glasgow Coma Scale,hyperglycemia,creatinine and smoker in logistic regression models. Depending on the factors entered into the model,either no effect or harm could be attributed to leech powder. Conclusions Based on the results of this study,no recommendations can be made on the use of leech powder in acute intracerebral hemorrhage. And properly randomized,controlled trials should be performed to come to a final conclusion.

[Objeetive] To develop the recombinant human bone morphogenetic protein-2(rhBMP-2)loaded amorphous calcium phosphate(ACP)delayed release nano-sized material,investigate its cytotoxicity of cell,and provide a reference for the experiment of composite material in vivo.[Method]The rhBMP-2/ACP delayed release nano-sized material were prepared by chemical wet method and cultured on rabbit bone marrow-derived mesenchymal stem cells(BMSCs)in vitro.Then the adhesion,proliferation,growth and functional expression of BMSCs were measured.[Result]Cytotoxicity test demonstrated that rhBMP-2/ACP delayed release nano-sized material had not affect the percentage of cell's proliferation with material-extracted liquid cultured with BMSCs and the cytotoxicity was graded zero.The adhesion,proliferation,configuration of the cells on the surface of this material were identical to the control group.[Conclusion]It was suggested that rhBMP-2/ACP delayed release nano-sized material might have good cellular biocompatibility,no cytotoxicity and not effected the normal functional expression of BMSCs in vitro.

[Objective]To study the biocompatibility and security of the recombinant human bone morphogenetic protein-2(rhBMP-2) loaded amorphous calcium phosphate(ACP) delayed release nano-sized material and investigate the feasibility of the clinical use as a kind of bone substitute in bone engineering.[Method]The in vitro hemolyzation,cytotoxicity,microkernel test in marrow smear,acute systemic toxicity,pyrogenicity,subcuticular stimulation reaction and short-term intramuscular implantation,as well as endosseous implantation were performed on the rhBMP-2/ACP delayed release nano-sized material.[Result]The material-extracted liquid induced no hemolyzation,no toxic effects of genetic,no pyrogenic reaction in rabbits,no cytotoxicity cultured in rabbit bone marrow-derived mesenchymal stem cells(BMSCs) in vitro and no acute toxicity in mice.The intramuscular implantation and endosseous implantation in rabbits induced no inflammatory reaction,tissue necrosis and the material was degraded gradually,fused organically with tissues.[Conclusion]The biocompatibility and security of the rhBMP-2/ACP delayed release nano-sized material could meet the requirements given in biological standards for implanted biomaterials ISO10993 and GB/T16886,suggesting that it could be a good bone substitution for the clinical trial.

In this paper, the mobile stereotactic radiosurgical system, a new therapeutant for interstitial radiotherapy, is introduced. Its principle and initial clinical applications are also summarized

Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

Objective To investigate the recovery of affected side kidney function after upper urinary tract obstruction was relieved Methods 78 patients who had been diagnosed with upper urinary tract obstruction were enrolled from January to December of 2015 in our hospital.All patients received the surgery to relieve the obstruction.GFR of the affected side kidney was done after one months of the surgery.The average age was(51.3 ± 12.8)years old.The reason of obstruction was upper urinary tract calculi (72 cases) and upper urinary tract stenosis (6 cases) respectively.All the patients received CT and ECT before and after operation.All the patients was divided into 3 groups by the decreased degree of affected side kidney function,including moderately decreased group [15 rnl/min ≤ GFR ＜ 30 ml/min,n =43,mean value =(23.1 ±5.0) mL/min],severely decreased group [7.5 rnL/min≤ GFR ＜ 15 ml/min,n =23,mean value =(11.2 ± 2.3) ml/min],and extreme severely decreased group [GFR ＜ 7.5 ml/min,n =12,mean value =(4.3 ± 2.9)ml/min].Linear correlation analysis was used to analysis the relationship analysis between the GFR value (pre-operation,post-operation) and the renal cortical thickness.The follow up time of extreme severely decreased group extended to 8-12 months.Results The GFR of moderate decreased group recovered to(30.6 ± 8.5) ml/min,regained averagely (7.56 ± 7.62) ml/min;the severely decreased group recovered to (13.1 ± 4.5) ml/min,regain (1.94 ± 3.38) ml/min.Extreme severely decreased group recovered to (11.1 ± 3.4) ml/min,regained averagely (6.75 ± 4.76) rnl/min,the GFR mean value after operation 8-12 months recovered to (12.7 ± 3.6) ml/min.All groups got significant recovery of kidney function of the affected side.The correlation coefficient between GFR value and the renal cortical thickness was 0.59 (before the surgery) and 0.70 (after the surgery) respectively (P ＜ 0.05).Conclusions Most of affected side kidneys got different degree of recovery.Obstruction influenced the accuracy of ECT at the time of evaluating the actual renal function before operation.The kidneys which had been supposed should be resected in presurgical evaluation could recover to the level of kidney reserve after the surgery.