BACKGROUND: A new material of porous carbonated hydroxyapatite cement (PCHC) is discovered using foaming technique.The new material characterizes original solidification and forms porous structure.OBJECTIVE: To investigate the biomechanical effect of PCHC on repairing cancellous bone defect.METHODS: Among 30 New Land rabbits, 25 ones were considered as surgery group, whose bilateral condyles of femur was used to establish bone defect model (5.5 mm diameter and 12 mm depth). PCHC was implanted into the left side, which was considered as the experimental group, and carbonated hydroxyapatite cement (CHC) was implanted into the right side, which was considered as the control group. Another 5 rabbits were used as normal mechanical control group. Both PCHC and CHC were dip in simulated body fluid (SBF) to test mechanical intension. PCHC and CHC were then implanted into muscles of back in the surgery group. Rabbits Were-sacrificed after 2, 4, 8, 12, and 16 weeks postoperatively. Mechanical analysis was tested following intra-bone and intramuscular implantation, and compressive strength was then tested following dipping into SBF.RESULTS AND CONCLUSION: PCHC: Intra-bone mechanical strength was lower at 2 weeks, the lowest at 4 weeks, but then closed to intension of normal cancellated bone at 8 weeks, higher than normal cancellated bone at 12 weeks, and recovered to the level of normal cancellated bone at 16 weeks. CHC: Intra-bone strength was higher than that of PCHC at 2 weeks, decreased at 4 weeks, gradually increased at 8, 12, and 16 weeks, but still lower than intension of normal cancallated bone. Compressive strength of both PCHC and CHC was not changed following dipping in SBF; however, compressive strength was changed remarkably following intramuscular implantation. The results demonstrated that PCHC characterized by immobilization in situ and mechanical supporting. Thus it could be used for one kind of bone substitute material to repair the bone defect.

AIM: RNAⅢ inhibiting peptide (RIP) has been previously proved to possess good histocompatibility and safety for preventing and curing staphylococcal infection, and this study evaluated histocompatibility of polyaiticglycolic acid/RIP (PLGA/RIP) sustained release microsphere. METHODS: The experiment was performed in the Orthopedic Institute, Pharmacologic Research Institute and Animal Experimental Center of General Hospital of Chinese PLA from October 2005 to October 2007.①Preparation of PLGA/RIP: The solid-phase synthesis (Fmoc) method was used to synthesize RIP from C end to N end, then the synthesized peptide was purified by the reverse phase high performance liquid chromatography, and composition was collected by means of ultraviolet absorption peak. The purified RIP was obtained after freezing and drying. Liquid-phase multiple emulsion method was used to synthesize PLGA/RIP microsphere of 50-70 ?m diameter.②Acute general toxicity test was studied in PLGA/RIP. Effect of PLGA/RIP on the cell proliferation was detected with cytotoxicity test by MTT method. Intramuscular implanting test was used to observe the irritation reaction of muscles by implantation materials. Sensitivity test was used to observe the sensitization of PLGA/RIP. Changes of animal's body temperature were determined with pyrogen test. RESULTS: ①Acute general toxicity test: Neither toxicosis reaction nor animal death was found after animals were injected with 100% and 50% eluents of PLGA/RIP peritoneally. Animal's body weight was not changed significantly.②Cytotoxiciity test by MTT method: The average proliferation rate of cell in two kinds of eluents exceeded 85% and cytotoxicity was graded in 1 rank, indicating no cytotoxicity.③Intramuscular implanting test: At 4 weeks after RIP and PLGA/RIP were implanted into the animals, there was not obvious synathresis, denaturation or necrosis in tissues. No inflammatory cell infiltration occurred around the materials. There had been the fibrous capsules around the materials.④Sensitivity test: Average primary irritation index of three groups were 0.38, 0.33 and 0.31 respectively. There was no significant difference among three groups.⑤Pyrogen test: Fervescence of each animal in the experiment was under 0.5 ℃, confirming that the materials had no pyrogenic characteristics. This was in coincidence with evaluation criterion of pyrogen test. CONCLUSION: PLGA/RIP has good histocompatibility and safety, without general toxic reaction, cytotoxicity, immunological rejection, hypersensitive response or pyrogenic characteristics.

Objective The aim of this study was to investigate the effect of ultrasonically activated hematoporphyrin on ultrastructure of ehrlich ascites tumor(EAT) cells and to evaluate the potential mechanism of action inducing this cytotoxicity. Methods EAT cells in vitro were exposed to ultrasound at 2^0?MHz and 1^5?W/cm+2 for 3?min in the presence or absence of hematoporphyrin.The changes of ultrastructure of sample preparation for different time were observed by transmission electron microscope. Results The degree of destruction of treated EAT cells was enhanced with the increasing of time for the sample preparation.The sites destroyed mainly involved cell membrane,mitochondrion,endoplasmic reticulum and cell nuclei.Furthermore,morphoiogical characters of ultrasound-activated hematoporphyrin induced apoptosis were observed on EAT cells.Conclusion The killing of tumor cells was ascribed mainly to the damage of ultrastructure induced by ultrasound in combined with hematoporphyrin,apoptosis was also induced during ultrasound and hematoporphyrin killing process.;

Objective: To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods: Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85αsiRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results: The expression of PI3Kp85 α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion: The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

[ ABSTRACT] AIM: To investigate the protective effect of autophagy on oxidized low density lipoprotein ( ox-LDL)-induced macrophage apoptosis and the underlying molecular mechanisms .METHODS:The RAW264.7 macropha-ges were pretreated with 3 mmol/L 3-methyladenine (3-MA), 1 μmol/L rapamycin (Rap) or 4 mmol/L 4-phenylbutyric acid ( PBA) respectively for 1 h and then treated with ox-LDL (100 mg/L) for 12 h.The cell viability and apoptosis were determined by MTT assay and flow cytometry with Annexin V-FITC/PI staining, respectively.The activities of lactate de-hydrogenase ( LDH) in the medium and caspase-3 in the cells were determined by detection kits .The protein levels of bec-lin-1 (a molecular marker of autophagy ), glucose-regulated protein 78 (GRP78, an endoplasmic reticulum stress marker) and C/EBP homologous protein ( CHOP, a key-signaling component of endoplasmic reticulum stress-induced apoptosis ) were examined by Western blot .Microtubule-associated protein 1 light chain 3 (LC3, another molecular marker of autoph-agy) was observed under laser scanning confocal microscope .RESULTS: Treatment of the RAW264.7 macrophages with ox-LDL at 100 mg/L for 12 h resulted in significant decrease in cell viability , and dramatic elevation in LDH leakage , cell apoptosis and caspase-3 activity, which were promoted by 3-MA (an autophagy inhibitor) and inhibited by Rap (an autoph-agy inducer ) .ox-LDL induced autophagy in the macrophages as assessed by beclin-1 upregulation and frequent granulation of LC3, which were inhibited by 3-MA and promoted by Rap.Interestingly, 3-MA enhanced, while Rap blocked, the CHOP upregulation induced by ox-LDL.Moreover , PBA ( endoplasmic reticulum stress inhibitor ) significantly inhibited ox-LDL-induced GRP78 upregulation and autophagy as determined by the attenuation of beclin-1 upregulation and frequent granula-tion of LC3.CONCLUSION: Endoplasmic reticulum stress mediates ox-LDL-induced autophagy in macrophages , and moderates activation of autophagy may protect macrophages from ox-LDL-induced apoptosis by inhibiting CHOP expression .

OBJECTIVETo evaluate the short term clinical results of minimally invasive total knee arthroplasty (MIS-TKA) through a quadriceps sparing (QS) approach compared with a standard TKA using a medial parapatellar exposure.

METHODSBetween March 2005 and March 2006, 26 consecutive unilateral primary MIS-TKA through a QS approach were performed. It was compared that the short term clinical results of this group with an age-matched and sex-match cohort of total knee arthroplasty done with a standard medial parapatellar approach. Length of incision, tourniquet time, blood loss, flexion degree, visual analog scale (VAS) score, Knee Society score, tibiofemoral angle and the ability to do a straight leg raising maneuver were evaluated respectively.

RESULTSThe mean follow-up time were 17 months (range 12-23 months). The average length of incision was (9.5+/-1.5) cm in the QS group and (14.0+/-2.3) cm in the standard group (P<0.05). Average tourniquet times were (83+/-16) minutes in the QS group and (55+/-11) minutes for the standard group (P<0.05). Postoperative tibiofemoral angles averaged (5.7+/-1.5) degrees valgus in the QS group knees and (6.0+/-1.4) degrees valgus in the standard group knees (P>0.05). The average VAS scores were significantly lower at the first, the third and the seventh postoperative days in the QS group compared with that of the standard group, and the difference was not significant after 6 weeks between two groups. Mean flexion at one week was greater for the QS group (107+/-12) degrees than that in the standard group (95+/-11) degrees. Improved ROM also was seen 6 and 12 weeks postoperatively in the QS group with significant difference. There were no differences in ROM between the two groups in 12 months. The ability to straight leg raised at one week postoperatively was better in the QS group (23/26, 88%) than that of the standard group (21/33, 64%). At the 6 weeks follow-up, the average knee score was (78+/-15) points in the QS group and (71+/-20) points for the standard group (P<0.05). Deep vein thrombosis were found in 1 case of the QS group and 3 cases for the standard group. One patient in the QS group developed superficial wound necrosis that healed uneventfully. There were no other complications such as deep infection, neurovascular injury and so on.

CONCLUSIONSThe MIS-TKA through a QS approach is associated with a more rapid functional recovery and improves range of motion. But higher specifications and more restrictive indications are of importance with the QS technique.

Arthroplasty, Replacement, Knee ; methods ; Follow-Up Studies ; Humans ; Minimally Invasive Surgical Procedures ; methods ; Quadriceps Muscle ; Treatment Outcome

OBJECTIVETo investigate the epidemiological features, genetic drift in the epitopes of hemagglutinin (HA) of the novel influenza A (H1N1) virus and oseltamivir-resistant variants characterized by H275Y and N295S mutations in children in Shanghai since the outbreak.

METHODBetween June 2009 and May 2012, a prospective surveillance study was carried out in Shanghainese children who attended the outpatient clinic of Children's Hospital of Fudan University for influenza-like illness. One-step real-time fluorescence quantitative RT-PCR was performed to detect seasonal influenza A and influenza B virus and the novel influenza A (H1N1) virus in the respiratory samples. Genetic drift from the vaccine strain in HA epitopes of the novel influenza H1N1 virus and the molecular markers associated with oseltamivir resistance in neuraminidase (NA) were analyzed.

RESULTOut of 3475 enrolled cases, the novel influenza A (H1N1) virus was confirmed virologically in 222 (6.4%) otherwise healthy children with 133 (59.9%) being boys and 89 (40.1%) girls. The median ages of children with the novel influenza A (H1N1) virus infection during the first wave from August 2009 to February 2010 and the second wave from December 2010 to February 2011 were 53.5 months and 32.0 months, respectively (Z = -4.601, P = 0.000); 119 (46.9%) had the close contact with persons suffering from fever or respiratory infection, of whom, 68 (57.1%) contacts were family members and 47 (39.5%) contacts were classmates. During the outbreak in 2009-2010 season, 66 (40.9%) were exposed to primary index cases, school students were the major exposure subjects, accounting for 50.0%. The nucleotide sequences of HA1 gene were highly homologous between the vaccine strain A/California/07/2009 and Shanghai circulating novel influenza A (H1N1) strains and only S83P mutation in epitope E of HA was detected inclusively in the circulating strains. The H275Y and N295S amino acid mutations associated with oseltamivir resistance were not found in the circulating novel influenza (H1N1) strains.

CONCLUSIONTwo major waves of the novel influenza A (H1N1) outbreaks occurred in Shanghainese children during 2009-2011. Institutional children were the major affected individuals during the 2009 pandemic wave. Households and schools were the main sites of transmission among children during influenza pandemic. Influenza vaccination should be enhanced in children and their close family contacts. The novel influenza A (H1N1) virus in Shanghai has not undergone significant genetic changes. Oseltamivir is effective for the treatment of the novel influenza A (H1N1) virus.

Adolescent ; Amino Acid Sequence ; Antiviral Agents ; pharmacology ; Child ; Child, Preschool ; China ; epidemiology ; Drug Resistance, Viral ; Female ; Hemagglutinins, Viral ; genetics ; Humans ; Infant ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; drug therapy ; epidemiology ; pathology ; virology ; Male ; Molecular Epidemiology ; Molecular Sequence Data ; Neuraminidase ; genetics ; Oseltamivir ; pharmacology ; Pandemics ; Viral Vaccines ; genetics ; immunology

OBJECTIVETo analyze the clinical and radiographical result of acetabular revisions with wire mesh, impacted irradiated frozen allografts and cemented cups.

METHODSFrom February 2006 to January 2009, a total of 20 patients with 21 acetabular revisions were performed with wire mesh, impacted irradiated frozen allografts and cemented cups. Eighteen cases (19 hips) were followed up. There were 5 hips in 4 males and 14 hips in 14 females. The average age of patients was 64.4 years (43 to 81 years). Acetabular bone defects were classified according to Paprosky classification. There were Paprosky II B in 4 hips, Paprosky II C in 8 hips, Paprosky IIIA in 5 hips and Paprosky IIIB in 2 hips. Wire mesh was used to converted segmental defects into cavity defects. Irradiated frozen allografts were impacted and cemented cup was inserted to complete the revision. Patients were followed up regularly with clinical and radiographical assessment. Harris score, migration and loosening of prosthesis grafts integration and complications were observed.

RESULTSThe average follow-up time was 22.4 months (12 - 48 months). Harris score improved from 42.5 points (31 - 56 points) pre-operation to 88.6 points (82 - 96 points) at the final follow up. Pain score was 14.4 point (10 - 20 point) before revision and 42.3 points (40 - 44 point) at the final follow up.

COMPLICATIONSthere was 1 infection and healing after debridement. One patient had weakness of quadriceps and returned to normal after 1 year. Greater trochanter fracture occurred in 1 patient. Cup migration and loosening were observed in 1 Paprosky IIIB patients. There was no cup migration more than 1 mm or change of abduction angle in the remaining 18 hips. Grafts incorporation defined as the presence of trabecular bone crossing the graft-host bond could also be seen in these 18 hips.

CONCLUSIONSImpacted bone grafting technique combined with wire mesh and cemented cup is an effective method for biological acetabular revision. Irradiated frozen allografts implanted with impaction bone grafting technique can integrate with the surrounding host bone.

Acetabulum ; surgery ; Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Bone Cements ; Bone Transplantation ; methods ; Female ; Follow-Up Studies ; Freezing ; Humans ; Male ; Middle Aged ; Prosthesis Failure ; Reoperation ; Surgical Mesh ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo study the clinical results of metal on metal hip resurfacing arthroplasty for developmental dysplasia patients.

METHODSFrom March 2005 to December 2006, 34 cases of developmental dysplasia patients (Crowe I, Crowe II) were attempted to have metal on metal hip resurfacing arthroplasty. There were 29 females (32 hips), 5 males (5 hips). The average age was 45 (26 - 57) years old. Radiographic and clinical evaluations were taken at 6 weeks, 3 months, 1 year and then once a year postoperatively. The average Harris score was 35 (25 - 44). Hip flexion was 101 degrees , abduction 24 degrees , adduction 15 degrees .

RESULTSThree patients were turned to total hip arthroplasty during operations. Thirty-one patients (34 hips) received hip resurfacing surgery. These 31 patients were followed for average 21.4 months (12 - 33 months). The average Harris score was 94 (82 - 100) at the latest follow-up, and there was statistical difference compared with the preoperative score (P < 0.01). Hip flexion increased to 133 degrees , abduction to 48 degrees , adduction to 26 degrees . No radiolucency line was found at both acetabular and femoral sides in all the patients. The average abduction angle of acetabular cup was 43 degrees (40 degrees - 53 degrees ), and the average stem shaft angle was 139 degrees (130 degrees - 145 degrees ).

CONCLUSIONSThe short term result is excellent. While the mid to long term results for hip resurfacing arthroplasty in developmental dysplasia patients are still looking forward, and the meticulous surgical technique and strict patient selection are the key of the good results.

Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo investigate the association between catecholamine-beta-adrenoceptor (beta-AR)-adenosine 3', 5'-monophosphate (cAMP) system and long-term prognosis in patients with chronic heart failure (CHF).

METHODSThe study population comprised 73 patients with CHF (EF: 23% +/- 10%) with a mean follow-up of 3.8 +/- 1.9 years. Plasma levels of norepinephrine (NE) were measured using high performance lipid chromatography, beta-adrenergic receptor density (Bmax) and the content of cAMP in peripheral lymphocytes were calculated using 3H-dihydroalpneolo as ligand and competitive immunoassay, respectively. Deaths due to cardiovascular events within the follow-up period were registered.

RESULTSThe total mortality was 64.7%, 57.4% of which was for cardiogenic (worsening heart failure: 32.4%; sudden death: 25.0%). In the cardiogenic death group, plasma levels of NE and epinephrine (E) (3.74 nmol/L +/- 0.09 nmol/L and 3.17 nmol/L +/- 1.0 nmol/L) and the contents of peripheral lymphocyte cAMP (3.64 pmol/mg protein +/- 1.4 pmol/mg protein) were significantly increased as compared with the survival group (2.68 nmol/L +/- 0.07 nmol/L, 2.41 nmol/L +/- 0.24 nmol/L and 2.73 pmol/mg protein +/- 0.9 pmol/mg protein, respectively, all P < 0.01). In the sudden death group, plasma levels of NE and E (5.01 nmol/L +/- 0.06 nmol/L and 4.13 nmol/L +/- 0.08 nmol/L) were significantly increased as compared with the worsening heart failure group (2.49 nmol/L +/- 0.07 nmol/L and 2.33 nmol/L +/- 0.8 nmol/L, all P < 0.001) and to the survival group (2.68 nmol/L +/- 0.07 nmol/L and 2.41 nmol/L +/- 0.14 nmol/L, all P < 0.01). The incidences of sudden death were 0%, 75%, and 100% (chi(2) = 16.018, P < 0.01) in patients with plasma NE < 2.5 nmol/L, NE 2.5 nmol/L - 4.5 nmol/L, and NE > 4.5 nmol/L, respectively. In the worsening heart failure group, the content of peripheral lymphocyte cAMP (4.46 pmol/mg protein +/- 0.18 pmol/mg protein) was significantly increased compared with the sudden death group (2.39 pmol/mg protein +/- 0.9 pmol/mg protein, P < 0.001) and to the survival group (2.73 pmol/mg protein +/- 1.1 pmol/mg protein, P < 0.001). The worsening heart failure death occurences were 5.0%, 72.2%, and 100% (chi(2) = 14.26, P < 0.01) in patients with a content of peripheral lymphocyte cAMP < 2.5 nmol/L, cAMP 2.5 nmol/L - 4.5 nmol/L, and cAMP > 4.5 nmol/L, respectively. Bmax in peripheral lymphocyte was not significantly different (P > 0.05) among the sudden death, worsening heart failure, and survival groups in CHF patients.

CONCLUSIONSPlasma levels of catecholamine increase significantly, and Bmax and the contents of cAMP in peripheral lymphocytes decrease significantly in patients with CHF. High plasma catecholamine levels may be associated with sudden death, and high intralymphocyte cAMP content may be associated with worsening heart failure in CHF patients.

Adult ; Aged ; Catecholamines ; blood ; Cyclic AMP ; blood ; Death, Sudden, Cardiac ; Female ; Heart Failure ; blood ; mortality ; Humans ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Receptors, Adrenergic, beta ; blood