BACKGROUND: Osteocytes are the main function cells in the femoral head. Therefore, the study of the apoptosis of osteocytes and the correlative stress genes has become the research focus for exploring the pathomechanism underlying avascular necrosis of femoral head. OBJECTIVE: To observe the expression changes of bcl-2 and bax genes during the development of traumatic avascular necrosis of femoral head in rabbits, and explore the pathomechanism underlying this disease. DESIGN, TIME AND SETTING: An animal observational experiment was performed at the Central Laboratory of Zhuhai Campus of Zunyi Medical College between December 2007 and September 2008. MATERIALS: A total of 35 New Zealand rabbits were randomly divided into 7 groups, namely, the control group and the experiment group which are subdivided into six ones at the time points of 3, 6, 12, 24, 48 and 96 hours postischemia respectively , with 5 animals in each group. METHODS: Rabbits in the experiment group were given an open surgery of interrupting the blood supply of their femoral heads to lead to avascular necrosis; rabbits in the control group only received skin incision and muscle-sparing ended in their joint capsules. Osteocytes, Bcl-2 and Bax proteins in femoral heads were determined and compared at different time points postischemia. MAIN OUTCOME MEASURES: Lacunae changes of osteocytes in femoral heads were detected with hematoxylin-eosin (HE) staining method, and the expression of Bcl-2 and Bax proteins with immunohistochemistry staining methods (ABC methods), after 3, 6, 12, 24, 48 and 96 hours of ischemia respectively. RESULTS: According to HE staining, ischemia of 12 hours or less resulted in no osteocyte lacunae change; after 48 hours of ischemia, parts of osteocytes and osteoblasts disappeared; at hour 96 postischemia, the percentage of empty osteocyte lacunae was higher obviously than that at hour 48 or that in the control group (P

[Objective]To investigate the methods of preparing acellular cartilaginous matrix(ACM) and constructing the tissue engineered cartilage with adipose-derived stem cells(ADSCs)-seeded acellular cartilaginous matrix in vitro.[Method]The ADSCs were isolated from adult New Zealand albino rabbits by collagenase,cultured and amplified in vitro.Fresh cartilage isolated from adult New Zealand albino rabbits were freeze-dried for twelve hours and then treated with Triton X-100,Dnase and RNase so as to obtain the ACM.After sterilized with ultra-violet,the ADSCs were seeded in the acellular cartilaginous matrix at a final density of 2?107/L and cultured in chondrogenic differentiation medium for two weeks in order to construct tissue engineered cartilage.Histology,immunohistochemistry and transmission electron microscope(TEM) were applied to examine the fresh ACM and tissue engineered cartilage.[Result]The test of hematoxylin-eosin(HE) staining and TEM showed no cellular structure in the ACM with only recesses left by removed cells.The ACM had suitable interval porosity and aperture size.After integrated with ADSCs,cells migrated into the ACM and adhered to the surface of material and grew well.After cultured in chondrogenic differentiation medium for two weeks,immunohistochemical staining of type Ⅱ collagen showed part of the cells in the material had enhanced expression of type Ⅱ collagen.[Conclusion]ACM can be used as scaffold material in cartilage tissue engineering.If it was seeded with ADSCs and cultured in chondrogenic differentiation medium,ACM can be used to construct cartilage tissue successfully.

BACKGROUND:Adipose-derived stem cel s and bone marrow mesenchymal stem cel s are used widely in cartilage tissue engineering, and there are many similarities in biological characteristics between two kinds of cel s. OBJECTIVE:To compare the chondrogenic potential of bone marrow mesenchymal stem cel s and adipose-derived stem cel s in vitro. METHODS:Adipose-derived stem cel s were isolated from the 3-month-old New Zealand white rabbits’ abdomen. Bilateral femurs of rabbits were obtained, and then the bone marrow mesenchymal stem cel s were separated with the adherence screening method. The growth curve of the passage 3 adipose-derived stem cel s and bone marrow mesenchymal stem cel s were drawn, and the doubling time of two kinds of cel s was compared. Then the passage 3 adipose-derived stem cel s and bone marrow mesenchymal stem cel s were treated with chondrogenic induction. After induced for 14 days, the adipose-derived stem cel s and bone marrow mesenchymal stem cel s were treated with toluidine blue staining and type Ⅱ immunohistochemistry staining respectively. RESULTS AND CONCLUSION:Primary bone marrow mesenchymal stem cel s showed aggregative growth, while the primary adipose-derived stem cel s were in single and scattered growth. The proliferation speed of adipose-derived stem cel s was faster than that of bone marrow mesenchymal stem cel s, while the doubling time of adipose-derived stem cel s was shorter than that of the bone marrow mesenchymal stem cel s. After chondrogenic induction for 14 days, both adipose-derived stem cel s and bone marrow mesenchymal stem cel s could express glycosaminoglycans and type Ⅱcol agen, and the expression level of type Ⅱ col agen in bone marrow mesenchymal stem cel s after chondrogenic induction was higher than that in the adipose-derived stem cel s. The in vitro proliferation of adipose-derived stem cel s and bone marrow mesenchymal stem cel s was rapid and stable, but the proliferative ability of adipose-derived stem cel s was faster than that of bone marrow mesenchymal stem cel s. When cultured in single layer, both adipose-derived stem cel s and bone marrow mesenchymal stem cel s could transform into chondrocytes under certain conditions, but bone marrow mesenchymal stem cel s seemed to be more potential than adipose-derived stem cel s.

The contents of protein, saccharides, ascorbic acid, and electrical potential, oxidation-reduction equivalent in 24 kinds of fresh vegetables and fruits were reported. The linear regression and correlation analysis were used to assess the relationship between health protection (nitrite eliminatin and antioxidation) and the nutrition characteristics of the materials. In a model system of simulated stomach condition, nitrite, one of the substrates for the formation of N-nitroso compounds, was efficiently eliminated by the active constituents of fresh vegetables and fruits. The correlation coefficient between the amount of eliminated nitrite and the antioxidative activity (inhibition rate of the formation of thiobarbituric acid-reactive substance) was 0.5115 (p0.05) , respectively. It was shown that the synthetic oxidation-reduction characteristic of nutrition constituents in fresh vegetables and fruits may be the natural cause of long life and health protection.

Objective To evaluate the protective effect of the angiotensin-converting enzyme inhibitor perindopril on diabetic glomerulopathy in rats with experimentally induced diabetes and study its possible mechanisms. Methods Changes of indexes, which include glomerular basement membrane thickness, mesangial matrix volume density, urinary protein excretion rate, plasma and renal tissue renin activity 、angiotensin Ⅱ concentration and renal angiotensinogen mRNA expression level in rats with experimentally induced diabetes, were observed. Results Angiotensin Ⅱ concentration increased in both the renal tissue and plasma in diabetic rats. Perindopril decreased proteinuria and delayed the progression of glomerular basement membrane thickening. However, it did not reduce the expansion of the mesangial matrix ( P

Objective To investigate the optimal dose of apocynin to protect severe acute pancreatitis (SAP) and SAP caused intestinal injury in rats. Methods A total of 53 SPF male Wistar rats were randomly allocated into five groups:sham operation group (SO group, n=10), SAP group (n=12), low-dose apocynin group (25 mg/kg,n=11), medium-dose apocynin group (50 mg/kg, n=10) and high-dose apocynin group (100 mg/kg,n=10). SAP model was prepared by retrograde infusing 5%sodium taurocholate (1 mL/kg) into biliopancreatic duct of rat. At thirty minutes before modeling, apocynin was injected into rat to intervention. The survival condition was recorded at 12 h after modeling, and blood samples were obtained for detecting serum amylase (AMY), alanine aminotransferase (ALT) and creatinine (Cr). Pancreatic and ileal tissue samples were obtained for HE staining and pathological examination. Results Two rats died in SAP group and one died in low-dose apocynin group. The quantity of ascites, the levels of AMY, ALT, Cr and pancreatic and intestinal pathologic scores were significantly increased in SAP group than those in SO group (P<0.05). Except the levels of Cr and intestinal pathologic score, there was no significant difference between low-dose apocynin group and SAP group. The quantity of ascites ascites, levels of AMY, Cr and pancreatic and intestinal pathologic scores were significantly lower in medium-dose and high-dose apocynin groups than those in SAP group (P<0.05). The levels of ALT and Cr were significantly higher in high-dose apocynin group than those of medium-dose apocynin group (P<0.05). Conclusion Apocynin improves SAP symptoms and reduces SAP caused intestinal injury in rats, which may be related to the inhibition of NOX activity, and 50 mg/kg of apocynin is the optimal dose.

BACKGROUND:Half-ring sulcated external fixator is a bone external fixation device.Its structure is relatively simple,with multi-plane fixed,and small occupying space.So far,there are few experimental reports about construction of tibial defect model by it in large animals.OBJECTIVE:To validate the practicality and repeatability of half-ring sulcated external fixator in establishing tibial defect models in goats.DESIGN,TIME AND SETTING:The randomized controlled observation was performed at the Animal Laboratory of the Third Military Medical University from March 2005 to February 2007.MATERIALS:Nine healthy adult(Animal Center of Third Military Medical University);Half-ring sulcated external fixator,made by Professor Li of the Third Military Medical University;Osseous pin,ψ2.5 mm,Shanghai Medicai Instruments Co.,Ltd.METHODS:After animals were anesthetized,two needles were threaded in the same plane of cancellous bone about 1.5-2.0 cm above the superior articular surface of the tibia,named group 1.The angle between the two needles was 40°.One needle was threaded at the place of 3.0 to 5.0 cm under the group 1 and parallel with optional needle of the group 1,named group 2.Needles of group 4 were threaded at the cancellous bone that was 2.0 cm from the joint surface of inferior extremity of the tibia.One needle was threaded at the place of 3.0 to 5.0 cm from group 4 and parallel with optional needle of the group 4.The included angle between these two needles of group 2 and group 3 was 60°.There were totally 6 needles.Tibia was fixed by half-ring sulcated external fixator.According to the length of the tibia from X-ray,tibia and periosteum were amputated by wire saw between the two needles in the middle and inferior segment of the tibia to make segmental bone and periosteal defects of 20 percent of tibia.MAIN OUTCOME MEASURES:①Postoperative general state of the animals and effect of half-ring sulcated external fixator.②Self-repair of bone defects by X-ray and Lane-Sandhu scoring standards.RESULTS:Of the 9 selected goats,1 died of postoperative infection,and 1 developed pinhole infection and supplemented.All the others survived and were included in final analysis.The goats were awake within 2-6 hours after surgery and able to stand up to eat.The loosening nuts were screwed up.No external fixation failure or loosening was found.No skin was necrotic.Injured limb could touch the ground in 2 days,but could not bear load until 2 weeks.The goats could walk in three weeks with a slight limp,then moved freely without claudication in four weeks after the operation.At the 5th week after operation,the radiographs showed little periosteal reaction.At the 10th week after operation,sclerosis was found in the fracture ends,and medullary cavity started to close.Fifteen weeks after operation,the medullary cavity closed and periosteal reaction did not aggravate.Inaddition.Lane score was 0 at any time point.No bone formation was found in the histological examination at 5,10 and 15 weeks after operation,and Lane score was 0 at any time point.CONCLUSION:The goat model established in goat tibia with 20%defects by the half-ring sulcated external fixator shows no bone healing after 15 weeks by X-ray or histological exanimation,indicating this is a practical and repetitive method to establish animal models of large segmental defect for bone tissue engineering.

BACKGROUND:There are reports concerning differentiation of adipose-derived mesenchymal stem cells(ADSCs)into chondrocytes using gene transfection technique.However,the transfection of adenovirus and adeno-associated virus into ADSCs is various.It is controversial whether adeno-associated virus(AAV)can transfect ADSCs.OBJECTIVE:To observe the enhanced green fluorescent protein(EGFP)expression following Ad5-EGFP and rAAV2-EGFP transfection into ADSCs,and investigate the cell proliferation ability following transfection.METHODS:ADSCs were isolated from the adipose tissue,which was from 6-month-old New Zealand albino rabbit back and neck by mechanical digestion and enzyme digestion,then ADSCs were cultured and amplified in vitro.ADSCs were infected with Ad5-EGFP and rAAV2-EGFP,and the EGFP expression was observed.A total of 2 μL sodium butyrate(1 mol/L)was added into the medium after rAAV2-EGFP transfection.MTT assay was used to detect the gene transfection effects on reproductive activity of ADSCs.RESULTS AND CONCLUSION:ADSCs isolated and cultured in vitro were flat,long-spindle and amplified stabry.The cell morphology was uniform.Many green fluorescent cells were observed in Ad5-EGFP and rAAV2-EGFP groups.Transfection efficiencies were about 88% and 83%.Adenovirus and adeno-associated virus vector can be transfected with ADSCs,and transfection efficiency is high.Adeno-associated virus needs sodium butyrate to increase its level of gene expression.

Objective To compare the effects of application of clavicular hook plate combined with wire anchors anatomical coracoclavicular ligament reconstruction and application of clavicular hook plate in the treatment of NeerII distal clavicle fracture and Tossy Ⅲtype~V acromioclavicular joint dislocation. Methods A retrospective analysis of the clinical data from June 2006 to June 2013. Total 73 cases patients suffered with Neer Ⅱtype distal clavicle fractures and Tossy Ⅲtype~V acromioclavicular joint dislocation. Of which , 41 cases were subjected to treatment with clavicular hook plate , 32 patients subjected to treatment of using clavicular hook plate combined with anchors .The incision length, operative time, postoperative complications, length of hospital stay and postoperative 1 month, 6 month shoulder VAS score of two groups were analyzed; the shoulder function of both groups after 1 month, 6 months were assessed by using Constant shoulder function assessment method. Results Surgical incision length and operational time between the two groups were statistically significant (P<0.05), while the amount of bleeding was not statistically significant. All patients were followed up . The two groups did not occur any complications such as loosening, decoupling, acromioclavicular joint dislocation and wound infections. Hospitalization time was 5~14 d (averaged 10d), no significant difference between two groups. 4 the shoulder Constant score and VAS scores showed no significant difference 1 months postoperation; 6 months after hook plate removed, VAS score and Constant shoulder score improved significantly in anchors hook plate group (P<0.05). Conclusion Anatomical coracoclavicular ligament reconstruction by application of hook plate combined with anchors is a good biomechanical model characterized with simple surgery , less trauma and good clinical outcomes , worthy of clinical application.

OBJECTIVE To investigate the effect of sodium ferulate (SF) on lipopolysaccharide (LPS)-induced preterm delivery and intra-uterine fetal death (IUFD). METHODS Pregnant Kunming mice were subcutaneously pretreated with SF (25 or 50 mg · kg-1) from gestational day (GD) 10 to GD 15 and with the single injection of LPS (150μg·kg-1, ip) on GD15.5. The incidence of preterm delivery and IUFD was observed. HE staining was used for uterine and placental histological evaluation. The levels of thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH) as well as the activities of glutathione S-transferase (GST) and glutathione peroxidase (GSH-Px) were detected in the maternal liver, placenta, and fetal liver using commercial kits. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) levels in amniotic fluid were evaluated by enzyme linked immunosorbent assay. RESULTS For LPS group, the incidence of preterm was 47.8%, delivery time was (17.5 ± 1.3) d, and the pups′survival rate was only 42.6%. Compared with LPS-treated group, SF 50 mg · kg-1 group showed a lower incidence of preterm (14.3%, P<0.01), longer gestational days (18.4 ± 0.5, P<0.05), and a higher pups′survival rate (75.6%, P<0.01). SF 50 mg · kg-1 restored the LPS-induced GSH both in the maternal and fatal liver (a tendency without statistical significance), GST activity〔(163±82) kU·g-1 protein vs (95±90) kU·g-1 protein, P<0.01)〕in the placenta, TBARS content〔(2.5±0.4)μmol·g-1 protein vs (3.1±0.6)μmol·g-1 protein, P<0.01〕in the fetal liver, and TNF-αlevel〔(11±8) ng·L-1 vs (20±8) ng·L-1, P<0.01〕in the amniotic fluid. SF also attenuated LPS-induced placental congestion and neutrophil infiltra?tion in the uterus. CONCLUSION SF may protect against LPS-induced preterm delivery and IUFD, and anti-oxidation as well as anti-inflammation may contribute to these effects.