Objective To investigate the correlation between nuclear factor(NF)-?B and inflammation of cell in adriamycin-induced nephritic rats,and the intervention of antisense oligonucleotide targeting NF-?B to adriamycin-induced nephritic rats by measuring the expression of NF-?B p65 in renal cortex and the secretions of interleukin-6(IL-6),tumor necrosis factor-?(TNF-?)in serum.Methods Thirty male SD rats were divided into 5 groups(6 rats in each group):group A(sham operation),group B(glomerulosclerosis),group C(sense oligonucleotide),group D(non-sense oligonucleotide),group E(antisense oligonucleotide).Glomerulosclerosis models were made for SD rats by unilateral nephrectomy and being injected with adriamycin into caudal vein.In the 8th week,various medicine applys to correspon-ding group for intervention.At the 4th day and 8th day after intervention,the excretion amounts of 24 hours urine protein were determined.Hematoxylin-Eosin(HE) staining was used to observe the renal pathological changes.Using image analysis system to calculate glomerulosclerosis index(GI).The expression of NF-?B p65 in renal cortex was measured by immunohistochemistry staining,the secretions of IL-6,TNF-? in serum were performed by means of enzyme-linked immunosorbnent assay,respectively.Results At different time points,the expressions of active-NF-?B p65,IL-6,TNF-? in group E were significantly lower than those in group B.The intervention effects of antisense oligonucleotides at the 8th day were stronger than that at the 4th day.Conclusions NF-?B is significantly correlated with glomerulosclerosis and inflammations of glomerular intrinsic cells.Using NF-?B antisense oligonucleotide to be injected into renal artery can block directly the translation of NF-?B gene,delay inflammations of glomerular intrinsic cells and the progressions of glomerulosclerosis.

OBJECTIVETo compare clinical outcomes of minimally invasive percutaneous plate osteosynthesis (MIPPO) in treating distal tibial comminuted fractures at early and delayed stage.

METHODSFrom January 2006 to January 2012,66 patients with distal tibial comminuted fractures were treated by MIPPO. All patients were divided into primary group and delayed group according to operation time. There were 31 patients in primary group, including 18 males and 13 females aged 21 to 57 years old with an average of (39.0 +/- 17.8), treated by MIPPO at primary stage,according to Tscherne soft tissue injury, 18 cases were grade I ,12 cases were grade II and 1 case were grade III. Thirty-five patients were treated by MIPPO at delayed stage, including 16 males and 19 females aged 24 to 55 years old with an average of (39.5 +/- 15.2), according to Tscherne soft tissue injury, 6 cases were grade I, 26 cases were grade II and 3 cases were grade III. Operation time, blood loss, hospital stay, fracture healing time and complications of two groups were recorded and observed, Lowa scoring of ankle joint were used to evaluated therapeutic effects at final following and AP and lateral X-rays were used to evaluated fracture reduction and alignment.

RESULTSAll patients were followed up, the time of following-up of primary group was (13.5 +/- 3.5) months, (15.2 +/- 3.8) months in delayed group, there was no significant meaning between two groups (t = 1.882, P = 0.064). There was no significant differences between two groups in operation time and blood loss (P > 0.05), but hospital stay in primary group was shorter than that of delayed group(P<0.05). There was no significant meaning between primary group (5.5 +/- 2.8) and delayed group (6.2 +/- 3.1) in fracture healing time (t = 0.958, P = 0.342); there was no significant meaning between primary group (87.6 +/- 6.8) and delayed group (89.6 +/- 5.2) in Lowa scores at final following-up (t = 1.351, P = 0.182). Two cases occurred postoperative superficial inflammatory reaction around fibular incision in primary group, 1 case occurred postoperative superficial inflammatory reaction around fibular incision and 1 case occurred delayed deep incision infection in delayed group at four months after operation. There was no significant differences in incidence of postoperative soft tissue complications between primary group (6.5%) and delayed group (5.7%) (t = 0.016, P = 0.900).

CONCLUSIONFor distal tibial comminuted fractures with grade I and II of Tscherne soft tissue injury, MIPPO at primary stage can not increase incidence of soft tissue complications, also can obtain the same clinical outcomes just like delayed MIPPO.

Adult ; Bone Plates ; Case-Control Studies ; China ; epidemiology ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; adverse effects ; methods ; Postoperative Complications ; epidemiology ; etiology ; Tibial Fractures ; surgery ; Time Factors ; Treatment Outcome ; Young Adult

Objective To evaluate the effecof bilateral ovariectomy combined with hormone injection on the bone mineral density and biomechanical property of sheep proximal femu.Method16 healthy adulsheep were divided into the sham operation group (n=8) and the experimengroup (n=8) randomly .Bilateral ovariewere only exposed in the sham operation group .The ex-perimengroup waperformed bilateral ovariectomy (OVX) and began to conducthe intramusculainjection of methylprednisolone (0 .45 mg · kg -1 · d-1 ) aftepostoperative 1 month fo10 month.The bone density (BD) of all sheep proximal femuwameas-ured before OVX and in postoperative 1 yea.The compression tesand the axial pullouteswere performed to evaluate biome-chanical property of postoperative 1 yeaproximal femu.ResultBD of proximal femubefore surgery had no statistically signifi-candifference between the two group,and which in the sham operation group had no statistically significandifference between before and aftesurgery (P＞0 .05) .BD of proximal femuin postoperative 1 yeain the experimengroup wasignificantly de-creased and significantly lowethan thain the sham operation group (P＜0 .05) .The maximal compression stresand the energy absorption value in the experimengroup were significantly lowethan those in the sham operation group with statistically signifi-candifferences(P＜0 .05);the maximal axial pulling force and the energy absorption value in the experimengroup were signifi-cantly lowethan those in the sham operation group with statistically significandifference (P>0 .05) .Conclusion The method of bilateral ovariectomy combined with hormone injection can significantly decrease BD and biomechanical intensity of sheep proximal femu.

Rheological properties of poloxamer 407 (brand named Pluronic F127) were examined by changing shear rate, temperature and the recovery properties of apparent viscosity after heating for several times. The results indicated that poloxamer 407 aqueous solution showed a Newtonian behavior at a low concentration while it might be a pseudoplastic fluid when the concentration reached a certain point. The thixotropy and the sol-gel transition temperature decreased with increasing the concentration (it could be an in situ gel at body temperature when the concentration of poloxamer 407 up to 15.25%). The results that obtained from the theological data would be useful in the application of poloxamer 407 such as in situ gel preparation.
Dose-Response Relationship, Drug ; Drug Compounding ; Drug Delivery Systems ; Excipients ; administration & dosage ; chemistry ; Poloxamer ; administration & dosage ; chemistry ; Rheology ; Shear Strength ; Solutions ; Temperature ; Viscosity ; Water

Acute-Phase Proteins ; metabolism ; Animals ; Female ; Hepatitis, Autoimmune ; immunology ; metabolism ; pathology ; Interleukin-17 ; metabolism ; Lipocalin-2 ; Lipocalins ; metabolism ; Liver ; pathology ; Mice ; Mice, Inbred BALB C ; Oncogene Proteins ; metabolism

Objective:To compare clinical outcomes of minimally invasive percutaneous plate osteosynthesis (MIPPO) in treating distal tibial comminuted fractures at early and delayed stage. Methods:From January 2006 to January 2012,66 pa-tients with distal tibial comminuted fractures were treated by MIPPO. All patients were divided into primary group and delayed group according to operation time. There were 31 patients in primary group ,including 18 males and 13 females aged 21 to 57 years old with an average of(39.0±17.8),treated by MIPPO at primary stage,according to Tscherne soft tissue injury,18 cases were gradeⅠ,12 cases were gradeⅡand 1 case were gradeⅢ. Thirty five patients were treated by MIPPO at delayed stage , including 16 males and 19 females aged 24 to 55 years old with an average of (39.5±15.2),according to Tscherne soft tissue injury,6 cases were gradeⅠ,26 cases were gradeⅡand 3 cases were gradeⅢ. Operation time,blood loss,hospital stay,frac-ture healing time and complications of two groups were recorded and observed ,Lowa scoring of ankle joint were used to evalu-ated therapeutic effects at final following and AP and lateral X rays were used to evaluated fracture reduction and alignment. Results:All patients were followed up,the time of following up of primary group was(13.5±3.5) months,(15.2±3.8) months in delayed group,there was no significant meaning between two groups (t=1.882,P=0.064 ). There was no significant differences between two groups in operation time and blood loss (P>0.05),but hospital stay in primary group was shorter than that of de-layed group(P<0.05). There was no significant meaning between primary group(5.5±2.8) and delayed group(6.2±3.1) in frac-ture healing time(t=0.958,P=0.342);there was no significant meaning between primary group(87.6±6.8) and delayed group (89.6±5.2) in Lowa scores at final following up(t=1.351,P=0.182). Two cases occurred postoperative superficial inflammatory reaction around fibular incision in primary group ,1 case occurred postoperative superficial inflammatory reaction around fibu-lar incision and 1 case occurred delayed deep incision infection in delayed group at four months after operation. There was no significant differences in incidence of postoperative soft tissue complications between primary group (6.5%) and delayed group (5.7%)(χ2=0.016,P=0.900). Conclusion:For distal tibial comminuted fractures with gradeⅠandⅡof Tscherne soft tissue in-jury,MIPPO at primary stage can not increase incidence of soft tissue complications ,also can obtain the same clinical outcomes just like delayed MIPPO.

OBJECTIVETo prepare ligustrazine (TMPZ) ocular sustained-release implant, and investigate its in vitro drug release, pharmacokinetics in rabbit vitreum and in vitro correlation.

METHODLigustrazine ocular sustained-release implants were prepared by micro-twin conical screw mixers with hot-melting extrusion method, with polyactic-co-glycolic acid (PLGA) as the matrix. HPLC was adopted to determine the concentration in vitreum after ligustrazine was implanted in rabbit eyes, in order to examine its in vivo sustained-release behavior, and study the correlation between in vitro and in vivo.

RESULTLigustrazine implants were prepared with a drug-loading rate between 10% and 30%, which was in conformity to the pharmacopoeia in terms of the content uniformity. Its in vitro release was in conformity to the zero-order release model. With PLGA 5050, 2. 5A as a vector, ligustrazine implants with a drug-loading rate of 30% could slowly release drug for more than 3 weeks, indicating a good correlation between in vitro and in vivo release.

CONCLUSIONLigustrazine ocular implants prepared with hot-melting extrusion method is practicable. Ligustrazine ocular implants release drug smoothly in rabbit vitreous vitreums, suggesting good sustained-release effect.

Animals ; Biological Availability ; Drug Implants ; Eye ; Female ; Male ; Polyglycolic Acid ; chemistry ; Pyrazines ; administration & dosage ; chemistry ; pharmacokinetics ; Rabbits ; Vitreous Body

The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.
Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Fibroblasts ; cytology ; physiology ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Mice ; Peptides ; chemistry ; Water ; chemistry

Without serum to provide adherent factors, CHO-dhfr- cells grow in suspension when cultured in serum-free medium. Although this offers advantages in some applications, in most production systems adherent cell growth is preferable. Gene transfection, clonal selection and amplification can be easier for adherent cells; the density of immobilized cells is often higher than those in suspension culture, which results in a higher protein productivity; washout of cells by perfused medium during continuous fermentation can be avoided; for high-throughput microplate assays, adherent cells are preferred to facilitate medium changes and cell washing. It has been proved that purified vitronectin alone was able to mediate attachment and spreading of CHO cells in serum-free medium. So we constructed a tricistronic expression vector expressing Igf-1, Vitronectin and Bel-2 at the same time. The vector was transfected into CHO-dhfr- cells and one clone, namely CHO-IVB2, expressing high level of the three proteins was screened out by Western blot. The cell line showed similar apoptosis-resistant and serum-independent properties to CHO-IB, an engineered cell line constructed before. When cultured in IMEM protein-free medium without any components supplemented, CHO-IVB can grow adherently. The viable cell numbers and growth rate of CHO-IVB were much higher than CHO-IB, making CHO-IVB an apoptosis-resistant host for production of recombinant proteins which can grow adherently in protein-free medium.
Animals ; Apoptosis ; CHO Cells ; physiology ; Cell Adhesion ; Cell Line ; Cell Proliferation ; Cricetinae ; Cricetulus ; Culture Media, Serum-Free ; Flow Cytometry ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; Vitronectin ; genetics

Mammalian cells are prone to apoptosis when cultured in large scale for production of biopharmaceuticals. And this will reduce production duration and result in high cost of production. Apoptosis is triggered by various factors, and delicately regulated by a set of genes. Bcl-2, a component integrated in mitochondria membrane, is an important member of these genes. By maintaining the integrity of mitochondria membrane, Bcl-2 keeps cytochrome C from releasing into cytoplasm, and thus blocks the activation of caspases, and subsequent onset of apoptosis. Over-expression of Bcl-2 has proven to be useful in blocking apoptosis in various cell lines, including CHO, hybridoma, myeloma, lymphoma and insect cells. Ammonia, a metabolite of cultured cells, however, showed apparent pro-apoptosis activity. In living cells, ammonia can be utilized by glutamine synthetase (GS) to synthesize glutamine, and thus lower the concentration of ammonia in medium, and its negative effects. Glutamine is essential to living cells. If not added into medium, glutamine can only be synthesized by GS, which makes GS a qualified selection marker. This marker can be used for gene amplification by adding into medium increased concentration of MSX, an inhibitor of GS. In this study, we over-expressed Bcl-2 using GS amplification in a recombinant CHO cell line stably expressing human interferon-beta. The modified cell line, with higher expression of Bcl-2 and lower production of ammonia, exhibited good anti-apoptosis quality and higher interferon-beta production in continuous culture.
Animals ; Apoptosis ; genetics ; physiology ; Biopharmaceutics ; CHO Cells ; cytology ; metabolism ; Cricetinae ; Cricetulus ; Glutamate-Ammonia Ligase ; genetics ; metabolism ; Interferon-beta ; metabolism ; Models, Genetic ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism