To evaluate the clinical therapeutic effect of the autologous bone marrow mesenchymal stem cells grafting integrating blood plasma transplantation in treatment of problematic nonunion. In March 2000, a 29-year-old man presented complaints of painful walking of left thigh after a velocity injury, was selected. Radiography revealed a left femoral shaft fracture, and the bone defect distance was 5 mm through X-ray examination before operation. Following a series of reposition, fixation, intramedullary nail fixation and twice autogenous lilac bone graft treatment, external fixator and autogenous lilac bone graft treatment, totally four times, the fracture was not healed. He come to Orthopaedic Surgery Department of Siping Central Hospital to accept the autologous bone marrow mesenchymal stem cells grafting integrating blood plasma transplantation in March 2008. Under the small C-arm X-ray perspective, 4 mL bone marrow-derived mesenchymal stem cells suspension integrating 10% autologous blood plasma was injected vertically into fracture site from the skin in front of the thigh with epidural needle, and the stem cells density was 1.8 × 107 cells/L. X-ray examination was performed though out-patient recheck to observe fracture healing. Two months after graft, radiography results showed fracture interspace reduced, left femoral shaft callus was continual and fracture lines blurred partly of the left femur; furthermore, left femoral shaft fracture lines blurred, and continuous bone callus formation of the left femur at 4 months after graft; The left femur achieved bone union within 7 months. The patient was returned to full weight bearing walking and good function with a fully healed femoral bone, without any fever or infection. Percutaneous transplantation of autologous bone marrow stem cells for treatment of problematic nonunion has the satisfactory result.

Objective To explore the variation tendency of serum thyroid hormone level in the elderly aged over 80 years.Methods The 602 healthy volunteers were divided into 3 groups by age:young group (20-59 years of age,n= 226),elderly group (60-79 years of age,n= 195),and advanced age group (80-102 years of age,n=181).Fasting blood of all persons was harvested,then the levels of serum total triiodothyroxine (TT3),total thyroxine (TT4),free tri-iodothyronine (FT3),free thyroxine (FT4),thyroid-stimulating hormone (TSH) and reverse tri-iodothyronine (rT3) were determined by chemistry luminescence technique and radioimmunoassay.Statistical analysis was made by the software SPSS 13.0.Results The levels of serum FT3 and TT3 were lower in elderly group than in young group (t=2.793,3.627,P=0.005,0.000).There were significant differences in the levels of serum TT3,TT4,FT3,TSH and rT3 between young group and advanced-age group (t =10.930,6.065,15.398,- 2.933,- 5.643,all P = 0.000),also between elderly group and advanced-age group (t= 8.382,4.298,11.573,-3.383,-5.148,all P＜0.001).The levels of serum FT3,TT3 and TT4 were negatively correlated with age (r=- 0.51,-0.39 and -0.25,respectively,all P＜0.01).And the levels of serum rT3 and TSH showed positive relationships with age (r=0.32,0.12,all P＜0.01).There were no relationships between the level of serum FT4 and age.The positive rate of serum TT3,TT4,FT3,FT4,TSH and rT3 concentration beyond the reference value was 0,0,13.8%,0,6.6% and 21% in advanced-age group,respectively.Conclusions The levels of serum thyroid hormone and thyroid-stimulating hormone change with age.The levels of FT3,rT3 and TSH change obviously in the elderly aged over 80 years.It could reduce the false positive rate in clinical practice if normal reference range for serum thyroid hormone levels in different aged elderly is established.

Objective To observe the mechanism of action of panax quinquefolium diolsaponins (PQDS) on anti-ventricular remodeling in rats. Methods The model of pressure-loading ventricular remodeling was established through the method of rat’s abdominal aorta deligation. The male Wistar rats were divided into 5 groups,including sham operation group,remodeling model group,Benazepril group,and low and high dose groups of PQDS. The rats were treated with PQDS (with dose of 50 and 100 mg?kg-1 i.g) for 6 weeks. The rats in sham operation group and remodeling model group were treated with normal sodium (with dose of 10 mL?kg-1?d-1 i.g) for 6 weeks. The rats in masculine medicine group were treated by Benazepril (with dose of 10 mg?kg-1 i.g) for 6 weeks. After 6 weeks of treatment,myocardial morphological parameters,the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in serum,the contents of prostacycline (PGI2),thromboxane A2 (TXA2),nitric oxide (NO) and angiotensinⅡ (AngⅡ) in plasma were determined. At the same time,the contents of endothelium (ET) in plasma and myocardium were also determined. Results Compared with remodeling model group,the ventricular weight and cardiac coefficient were decreased significantly in the rats treated with PQDS (P0.05). Conclusion PQDS has protective effects on ventricular remodeling through increasing anti-oxidase activity,reducing the damage of free radicals and vasoactive substance on myocardium and correcting disequilibrium of PGI2/TXA2 in ventricular remodeling etc.

Objective To discuss and analyze the issue in the catheter directed thrombolysis(CDT) treatment of acute lower extremity deep venous thrombosis.Methods The clinical data of 182 cases of acute lower extremity deep venous thrombosis from January 2011 to January 2013 which were treated by CDT were retrospectively analyzed,and the issues in these treatment were discussed and analyzed.Results The effect were classified into three degrees.Thrombosis clearance exceed 95 percent belonged to the first degree,among 50 percent and 95 percent belonged to the second degree,and below 50 percent belonged to the third degree.In this study,the respective number of each degree were 93/68/21.Sixty-one cases combined iliac venous compression syndrome were treated by stent.Twenty-two cases were treated with inferior vena cava filter.All the patients were followed up 10 months,148 cases of them maintained patency.Conclusions The effect of CDT treatment to acute lower extremity deep venous thrombosis is remarkable,meanwhile the patency is improved and the deep vein thrombosis were decreased through this method.But the issues of access,filter and iliac venous compression syndrome should also be considered.

This study examined the effect of insulin on the expression of very low density lipoprotein receptor (VLDLR) subtypes of SGC7901 cells and discussed its biological implication. In vitro, moderately or poorly-differentiated human gastric adenocarcinoma cell line SGC7901 was incubated with insulin for different lengths of time, and then the expression of protein and RNA level in VLDLR subtypes were detected by Western blotting and real-time PCR, respectively. The results showed that, at certain time interval, insulin could down-regulate expression of type I VLDLR and up-regulate the expression of type II VLDLR in SGC7901 cells, at both protein and RNA level. We are led to conclude that insulin serves as a regulator in maintaining the balance between glucose and lipid metabolism in vivo, possibly through its effect on the differential expression of VLDLR subtypes.

In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.

To explore the intracellular signal pathways for beta-VLDL induced very low density lipoprotein receptor (VLDL-R) transcription up-regulation and their effects on lipid accumulation in macrophages, Western Blot was used to examine phosphorylated ERK1/2 protein and regulated effects by different singal kinase inhibitants. It was found that beta-VLDL induced an increase in ERK1/2 activity in a protein kinase C (PKC)-dependent manner in murine RAW264.7 macrophages. By using different protein kinases inhibitors or activators, it was observed that the effect of beta-VLDL induced VLDL receptor transcription, which was monitored by RT-PCR analysis of VLDL receptor mRNA, was not affected by the inhibitor of p38 kinase and cAMP analog, but extremely abolished by pretreating cells with PD98059, an inhibitor of ERK and GF 109203X, an inhibitor of PKC. These results demonstrated that the PKC-ERK1/2 cascade is the essential signaling pathway by which beta-VLDL activated VLDL-R mRNA expression. Inhibition of the ERK1/2 signaling cascade resulted in suppression of the cellular lipid accumulation induced by beta-VLDL in macrophages.
Cells, Cultured ; Lipoproteins, VLDL ; metabolism ; Macrophages ; cytology ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; physiology ; Mitogen-Activated Protein Kinase 3 ; metabolism ; physiology ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Receptors, LDL ; biosynthesis ; genetics ; Signal Transduction ; Transcription Factors ; metabolism ; Transcription, Genetic ; Up-Regulation

In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column. The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully.

In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni+-nitrilotriacetic acid (Ni+-NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni+-NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.

This study examined the effect of insulin on the expression of very low density lipoprotein receptor (VLDLR) subtypes of SGC7901 cells and discussed its biological implication. In vitro,moderately or poorly-differentiated human gastric adenocarcinoma cell line SGC7901 was incubated with insulin for different lengths of time, and then the expression of protein and RNA level in VLDLR subtypes were detected by Western blotting and real-time PCR, respectively. The results showed that, at certain time interval, insulin could down-regulate expression of type Ⅰ VLDLR and up-regulate the expression of type Ⅱ VLDLR in SGC7901 cells, at both protein and RNA level.We are led to conclude that insulin serves as a regulator in maintaining the balance between glucose and lipid metabolism in vivo, possibly through its effect on the differential expression of VLDLR subtypes.