OBJECTIVETo investigate the short-term clinical outcome of unicompartmental knee arthroplasty for the treatment of spontaneous osteonecrosis of the knee.

METHODSFrom September 2013 to April 2014,5 patients with spontaneous osteonecrosis of the knee underwent unicompartmental knee arthroplasty, included 3 males and 2 females, aged from 65 to 80 years old with an average of 74 years. The courses of disease was from 1 to 6 years with the mean of 3 years. According to the radiographic staging criteria of Koshino, 1 case was stage II, 2 cases were stage III, 2 cases were stage IV. Clinical effects were assessed by VAS score, HSS score, and knee range of motion, tibiofemoral angle before and after operation.

RESULTSAll the patients were followed up from 6 to 7 months with an average of 6.4 months. All incisions obtained primary healing, and there were no complications such as infection, thrombosis, fracture of lower limbs. All 5 patients' pain relieved and their knee function improved significantly after operation, but knee range of motion had no obviously improved. Postoperative HSS scores, VAS scores, tibiofemoral angle were significantly improved than that of preoperative.

CONCLUSIONThe short-term effect of unicompartmental knee arthroplasty in treating spontaneous osteonecrosis of the knee is satisfactory.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Joint Diseases ; physiopathology ; surgery ; Knee Joint ; Male ; Osteonecrosis ; physiopathology ; surgery ; Range of Motion, Articular

Objective:To investigate the effect of astragaloside IV (AS-IV) on the secretion of stromal cell-derived factor-1α (SDF-1α) and CXC chemokine receptor 4 (CXCR4) by high glucose injured human umbilical vein endothelial cells (HUVECs), so as to lay a foundation for further study on AS-IV improving angiogenesis by regulating SDF-1 α/CXCR4 axis of endothelial cells.Methods:HUVECs were isolated and cultured from the umbilical vein of full-term healthy newborns and identified by von Willebrand factor (vWF) combined with 4-diamino-2-phenylindole (DAPI) nuclear staining. The obtained HUVECs was cultured in EGM-2 medium with 30 mmol/L glucose for 120 h to obtain high glucose damaged HUVECs. After intervention with different concentration gradients (25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) AS-IV for 72 hours, the contents of SDF-1α and CXCR4 were detected by enzyme linked immunosorbent assay (ELISA) method to determine the best concentration of AS-IV. The supernatant of damaged HUVECs were collected at 6, 12, 24, 48 and 72 hours after intervention with the best concentration of AS-IV, and the contents of SDF-1α and CXCR4 were detected by ELISA method to determine the best action time of AS-IV. The damaged HUVECs was randomly divided into experimental group and control group, and the blank group was set up at the same time. The experimental group was treated with the best concentration of AS-IV and the best time, the control group and the blank group were treated with the same volume of phosphate buffered saline (PBS) solution, and the contents of SDF-1α and CXCR4 in each group were detected by ELISA method.Results:The vWF factor on the cell membrane was green fluorescence, and the nucleus was blue after DAPI staining. When the fusion image showed green fluorescence, HUVECs were identified by blue fluorescence. The expression of SDF-1α in damaged HUVECs was the best when treated with AS-IV of 100 mg/L for 24 hours (1 642.87 pg/ml), and the expression of CXCR4 in damaged HUVECs was the best when treated with AS-IV of 50 mg/L for 48 hours (8.44 ng/ml). Compared with the control group, the contents of SDF-1α and CXCR4 in the experimental group were significantly increased, and the difference was statistically significant ( P<0.05). While the contents of SDF-1α and CXCR4 in the experiment group were slightly less than those in the blank group and there was no statistically significant difference ( P>0.05). Conclusions:AS-IV can promote the expression of SDF-1α and CXCR4 in HUVECs damaged by high glucose to return to normal physiological level, so as to play the role of vascular repair and neovascularization.

OBJECTIVETo explore the clinical value of Lens culinaris agglutinin-reactive alpha-fetoprotein detected by microspincolum method for judgment of treatment response in patients with hepatocellular carcinoma undergoing transcatheter arterial chemoembolization.

METHODSTwenty eight patients with HCC undergoing TACE follow-up in hospital were recruited. AFP and AFP-L3 were measured in all the patients before and after TACE, and correlations were analyzed between AFP-L3% and response to treatment.

RESULTSAmong the twenty eight patients with HCC undergoing TACE, 8 out 11 case in AFP-L3% descent group had well treatment response, 5 out 17 case in AFP-L3% elevated group had well treatment response(Chi2 = 4. 858, P < 0. 05).

CONCLUSIONSThe detection of AFP-L3 by microspincolum method is useful to judgment of treatment response in patients with HCC undergoing TACE.

Adult ; Aged ; Carcinoma, Hepatocellular ; chemistry ; therapy ; Chemoembolization, Therapeutic ; methods ; Female ; Humans ; Liver Neoplasms ; chemistry ; therapy ; Male ; Middle Aged ; Treatment Outcome ; alpha-Fetoproteins ; analysis

OBJECTIVETo evaluate indications and clinical results of total hip arthroplasties for degenerative hips with history of infection.

METHODSSeven cases of degenerative hip with history of infection underwent primary total hip arthroplasties, which involved 5 males and 2 females, with an average age of 45.8 years (range, 30 to 65 years). The quiescent period of infection were more than 10 years in all hips. According to Kim classification, 3 cases were of type I, and 4 of type II. The method to exclude active infection at the site of degenerative hips preoperatively was combination of physical examination, erythrocyte sedimentation rate and C-reactive protein level. The lateral incision was adopted in all cases, and all prosthesis were cementless. The clinical results of affected hips were assessed according to Harris hip score.

RESULTSThe follow-up was performed with the mean duration of 33.5 months (range, 21 to 44 months). No recurrence of infection, damage of nerve function or deep vein thrombosis of lower extremities occurred in all cases. The mean Harris hip scores improved from 44.5 points preoperatively to 84 points at the latest follow-up. No aseptic loosening of prosthesis or periprosthetic osteolysis were found at the latest follow-up.

CONCLUSIONTotal hip arthroplasties has good short term results for degenerative hips with history of infection. It is important to select indicated cases and rule out the possibility of active infection.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Bone Diseases, Infectious ; complications ; Female ; Follow-Up Studies ; Hip ; diagnostic imaging ; pathology ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Radiography

Objective To evaluate the feasibility and safety of a delivery pathway for the performance of percutaneous left atrial appendage (LAA) occlusion in experimental canine models. Methods Transseptal puncture was performed via femoral vein approach under fluoroscopic and angiographic guidance in 12 experimental dogs. A pigtail catheter was advanced into the left atrium (LA), which was followed by LA angiography. The diameters of the neck of LAA were measured on LAA angiogram obtained in appropriate projection. After the delivery sheath was advanced along the wire into LA, a pigtail catheter was inserted into the ostium of the LAA and the sheath was then advanced over the pigtail into the LAA. LAA angiography was then performed through the delivery sheath to confirm the position of the delivery sheath. One hour after the procedure both electrocardiography (ECG) and transthoracic echocardiography (TTE) were carried out in five dogs to check the results, immediately after which the five dogs were sacrificed to macroscopically observe the damages of the puncture site of inter-atrial septum as well as inside the LA and LAA. One hour and 2 weeks after the procedure TTE was conducted in the remaining 7 dogs and these dogs were followed up for one month. Results One dog died of pericardial tamponade during the operation. In 8 dogs the LAA was clearly displayed in the projection position of right anterior oblique (RAO) 30°/cranial (CRA) 20°,while in 3 dogs the LAA was well visualized in the projection position of RAO 30° , and in one dog in the projection position of RAO 30°/caudal (CAU) 20°. The diameter of LAA neck was (13.6 ± 5.2) mm. The delivery sheath was safely advanced into the LAA along the pigtail catheter in all dogs, and no air embolism, thrombus or pericardial tamponade occurred. Hematoma at puncture point of groin occurred in 2 dogs, which was absorbed through pressure dressing. Macroscopic examination of the heart performed immediately after the operation showed that no bloody pericardial effusion was found, and mild hematoma at posterior wall of LA was seen in one dog and mild damage of the upper-margin intima of LAA was noted in 2 dogs. The mean fluoroscopy time was (10.1 ± 2.5) minutes and the mean operation time was (58 ± 12) minutes. TEE showed no pericardial effusion 2 weeks after the procedure. During the follow-up period of one month no sudden death, stroke or infection occurred. Conclusion This method of placing the delivery sheath into the LAA is clinically safe and effective, and it can reliably establish a pathway to advance the LAA occluder into LAA.

OBJECTIVETo observe the effect of the supernatant of heat injured keratinocytes (KC) on biological behavior of the dermal fibroblasts (Fb).

METHODSHuman dermal Fb were isolated and cultured. A model of heat injured KC (HaCaT) was reproduced in vitro. Supernatant of normal KC and the supernatant of KC culture 12 hours after heat injury were collected and diluted with non-serum DMEM in 1:1 volume ratio to make normal KC conditioned medium (NKCM) and heat injury KC conditioned medium (HKCM) respectively. Fb was respectively treated with non-serum DMEM and 2 kinds of conditioned medium. (1) The proliferation of Fb was detected with MTT method at post culture hour (PCH) 12, 24, 36, 48. (2) The apoptosis of Fb was determined by flow cytometry at PCH 12 (Fb were heat injured in advance; Fb without heat treatment was used as control). (3) At PCH 24, expression of a-SMA in Fb cytoplasm was determined with immunofluorescence method; expression of a-SMA mRNA in Fb was determined with real-time quantified PCR. Data were processed with one-way analysis of variance, and pairwise comparison among groups with LSD-t test.

RESULTS(1) The proliferation of Fb: the absorbance value of Fb cultured with HKCM at PCH 12, 24, 36, 48 was respectively higher than that of Fb cultured with non-serum DMEM (with t value respectively 1.89, 2.35, 2.02, 1.94, and P values all below 0.01). There were significant statistical differences between the absorbance values of Fb cultured with HKCM and those of Fb cultured with NKCM at PCH 12, 24, and 48 (at PCH 12, t = 1.83, P < 0.01; at PCH 24, t = 2.91, P < 0.05; at PCH 48, t = 1.83, P < 0.05). (2) Apoptosis of Fb cultured with HKCM was diminished as compared with that of Fb cultured with NKCM and of Fb without treatment (t = 3.31, P < 0.05; t = 1.47, P < 0.01). (3) The expression of alpha-SMA (red fluorescence) in Fb cultured with non-serum DMEM or NKCM was less as seen under fluorescence scope, and it was obviously increased in Fb cultured with HKCM. (4) The relative expression amount of alpha-SMA mRNA in Fb cultured with HKCM was 1.32 +/- 0.06, which was higher than that both in Fb cultured with NKCM (1.14 +/- 0.07, t = 2.51, P < 0.05) and in Fb cultured with non-serum DMEM (1.00 +/- 0.09, t = 1.77, P < 0.05).

CONCLUSIONSThe supernatant of KC 12 hours after heat injury can obviously promote the proliferation of Fb, inhibit its apoptosis and accelerate transdifferentiation of Fb to myofibroblasts.

Actins ; metabolism ; Apoptosis ; Cell Differentiation ; Cells, Cultured ; Culture Media, Conditioned ; pharmacology ; Fibroblasts ; cytology ; drug effects ; metabolism ; Flow Cytometry ; Heat Stress Disorders ; Hot Temperature ; adverse effects ; Humans ; Keratinocytes ; cytology ; RNA, Messenger ; genetics

OBJECTIVE: To determine the effect of activation of specific anti-tumor cytotoxic T lymphocytes (CTL) and the ability of cross-presentation in vitro by fusion of HLA-A2+ human dendritic cells (DCs) with HLA-A2- melanoma cells. METHODS: The HLA-A2+ human dendritic cells and HLA-A2- melanoma cells were fused by PEG and were cultivated in complete RPMI1640 media containing FCS (10%) and GM-CSF for 24-48 h, and then co-cultured fusion cells with Melan-A specific T cells. HLA-A2- melanoma cells were negative control,While T2 cells and DC+Pts were positive control. The activation of anti-tumor CTL elicited by the fusion cells was detected by intracellular cytokine staining. RESULTS: The immature DC could express CD80, CD83, CD86, HLA-DR, and HLA-ABC,but the mature DC induced by TNF-alpha, PGE-2, and CD40L further highly expressed above molecules. The rate of specific CTL cells primed by the fusion cells was 16.72%+/-4.26%, negative control was 0.21%+/-1.84%,and positive control was 28.60%+/-5.67%. The CTL from vaccine by fusing DC and LAR6 induced lysis of HLA-A2+ LAR1 cells. CONCLUSION The HLA-A2 restricted specific anti-tumor CTL can be induced in vitro by fusion of HLA-A2+ human dendritic cells with HLA-A2- melanoma cells.
Antigen Presentation ; immunology ; Antigens, Neoplasm ; immunology ; Cancer Vaccines ; immunology ; Cell Fusion ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; HLA-A2 Antigen ; immunology ; Humans ; MART-1 Antigen ; immunology ; Melanoma ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo reproduce a model of heat injured KC in vitro and explore its apoptosis rate of KC due to heat injury at different temperature.

METHODSHuman KCs were cultured in vitro, and they were incubated at 37, 41, 43, 45, 48, and 51 degrees C respectively for 10 mins in water bath. Trypan blue staining and Hoechst 33258 fluorescence staining were used respectively to determine necrosis and apoptosis of KC. Rates of apoptosis and necrosis of KC were analyzed quantitatively by flow cytometer. The proliferation activity of KC after heat injury was detected by MTT test.

RESULTSThe results of trypan blue staining, Hoechst 33258 fluorescence staining, and flow cytometer demonstrated that number of apoptotic and necrotic KC increased gradually along with a rise of water bath temperature. The rates of apoptosis and necrosis of KC were respectively (12.3 +/- 3.2)% and (14.1 +/- 1.6)% at 45 degrees C, (27.7 +/- 5.1)% and (58.0 +/- 4.2)% at 48 degrees C. Rate of KC necrosis reached up to (83.0 +/- 5.3)% at 51 degrees C. Inhibition of KC growth reached a stationary phase when the injurious temperature reached 45 degrees C as observed with MTT test.

CONCLUSIONSHeat injury can induce apoptosis and growth inhibition of KC in vitro. Incubating KC at 45 degrees C for 10 mins is a good condition to reproduce a model of heat injured KC in vitro. This model may be used to study the biological character and apoptosis of KC after burn injury.

Apoptosis ; Burns ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Flow Cytometry ; Hot Temperature ; Humans ; Keratinocytes ; cytology

This study was aimed to investigate the expression and role of 14-3-3ζ in the AML cell lines: sensitive HL-60 and drug-resistant HL-60/VCR cells. Semi-quantitative RT-PCR and Western blot were respectively used to examine the expression of mdr1 mRNA and Pgp in AML cell lines to validate the results of microarray. Western blot was performed to investigate the expression of Pgp, 14-3-3ζ, and anti-apoptosis protein BCL-2, MCL-1 proteins. Immunofluorescence assay was used to detect the subcellular location of 14-3-3ζ protein in HL-60 and HL-60/VCR cells by laser scanning confocal microscopy. Transduction with siRNA was used to silence 14-3-3ζ in AML cell lines. Cell count method and flow cytometry of cell cycle were used to analyze the changes of growth of AML cells. The results found that mdr1 mRNA and Pgp did not expressed in HL-60 cells, but significantly overexpressed in HL-60/VCR cells. Except 14-3-3σ, the expression of other subtypes of 14-3-3 was higher in HL-60/VCR cells than that in HL-60 cells, especially 14-3-3ζ. The higher expression of 14-3-3ζ, BCL-2, MCL-1 protein was observed in HL-60/VCR cells than that in HL-60 cells. These results were same results from gene chip. It was also noticed that 14-3-3ζ was located in the cytoplasma and nuclei of AML cell lines, especially over-expressed in HL-60/VCR cells. Furthermore, suppression of 14-3-3ζ by RNA interference resulted in inhibition of the proliferation of AML cells with decreased protein expression of BCL-2 and MCL-1, especially in HL-60/VCR cells. It is concluded that 14-3-3ζ plays an important role in proliferation of AML cells and associates with BCL-2 and MCL-1 expression. These results suggested that development of therapy targeting 14-3-3ζ may provide novel, effective strategies for refractory and relapsed AML.
14-3-3 Proteins ; metabolism ; ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Apoptosis ; Cell Proliferation ; HL-60 Cells ; metabolism ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

OBJECTIVEIn order to investigate the positive rate of streptococcus suis type 2 and the genes of their suilysin (sly), extracellular protein (epf) and muramidasa-released protein ( mrp) and to understand the antibiotic susceptibility of S. suis type 2.

METHODSS. suis type 2, isolated from slaughtered healthy pig's tonsil in 10 county area of Guangxi, were identified by Multiplex PCR, and the genes of their sly, epf, mrp and the antimicrobial sensitivity analysis were performed.

RESULTS1105 strains of Streptococcus including 667 strains of S. suis and 33 strains of S. suis type 2 were detected from 1179 samples. In these S. suis type 2 strains, there were 22 strains of sly + mrp + epf+ type,1 strain of sly + mrp + epf - type, 2 strains of sly - mrp + epf + type, 7 strains of sly - mrp + epf - type and 1 strain of sly - mrp - epf- type. When these strains were subjected to be tested with penicillin, eritrocina, vacocin, gentamycin, specti-nomysin, enraxacin, ciprofloxaxin, cephalothin VI, sulfadiazine sodium, cyantin, mycifradin, amikacin and achromcin, some were found to be resistant to but most strains were susceptible to cephalothin VI, penicillin and enraxacin. There were 31, 29 and 27 strains over medium sensitivity, respectively, but 28 and 27 resistant strains to amikacin and achromcin were found.

CONCLUSIONThe positive rate of S. suis type 2 in clinical healthy pigs was low (2.8%) and did not show obvious difference between the counties with or without a history of S. suis infection. All the isolated strains were susceptible to cephalothin VI, but most strains were virulent.

Animals ; Anti-Bacterial Agents ; pharmacology ; Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; Drug Resistance, Bacterial ; genetics ; Hemolysin Proteins ; genetics ; Microbial Sensitivity Tests ; Molecular Epidemiology ; methods ; Polymerase Chain Reaction ; Streptococcal Infections ; epidemiology ; genetics ; microbiology ; Streptococcus suis ; drug effects ; genetics ; pathogenicity ; Swine ; Swine Diseases ; epidemiology ; genetics ; microbiology