OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To observe efifcacy and safety of catheter ablation for atrial ifbrillation (AF) occurring after surgical valve replacement in patients with rheumatic heart disease (RHD). Methods A total of 23 RHD patients with atrial ifbrillation after surgical valve replacement were enrolled in this study from 2008 to 2013. The clinical characteristics, ablation strategies and successful rate were investigated. Results All the cases included 8 males and 15 females (age, 51.0 ± 9.2 years). Valves replaced were isolated mitral valves (13/23, 56.5%) and multiple valves (10/23, 43.5%). Postoperative AF after cardiac surgery was paroxysmal in 14 patients (60.9%) and nonparoxysmal in 9 cases. Nine patients (39.1%) was in sinus rhythm before cardiac surgery, 4 in paroxysmal AF and 10 in non-paroxysmal AF. The mean interval between the catheter ablation AF and the surgical intervention was (6.9±5.8) years. The postoperative AF duration was (3.1±3.2) years, left and right atrial diameters were (44.1±5.9) mm and (48.1±9.0) mm respectively, left ventricular ejection fraction was 64.0%±8.3%, the mean ablation procedure duration was (156.8±46.6) min, and lfuoroscopy exposure averaged (27.3±11.2) min. Standard pulmonary vein isolation was performed in all cases by using ipsilateral circumferential ablation technique. Additional ablation, including complex fractionated atrial electrograms, mitral and tricuspid isthmus, and left atrial roof, was applied in most of the cases. After a mean follow-up of (29.7±21.2) months (median, 24 months), 60.9%of the patients remained free of AF, 1 died, and 2 lost to follow-up. Conclusions Catheter ablation for AF is effective and safe in patients with RHD after surgical valve replacement. Stepwise ablation strategy may be better for these patients.

Background Neovascular glaucoma (NVG) is a serious ocular disease which may cause blindness.The primary pathogenesis of NVG is ischemic retinopathy derived by central retinal vein occlusion (CRVO) and diabetic retinopathy (DR).Clinical characteristics of NVG are variable based on the difference of primary diseases,such as CRVO and DR.However,there is a few studies regarding the diffcrcnces of NVG initiated by CRVO and DR.Objective This study was to compare the clinical characteristics in NVG patients secondary to CRVO and DR.Methods A series case observational study was carried out in Hiserve Hospital of Qingdao University from January 2009 to June 2012.Twenty-nine eyes of 27 patients with NVG caused by CRVO (10 eyes of 10 patients) and DR (19 eyes of 17 patients) were included.The history of underlying diseases,course of NVG,intraocular pressure(IOP),fundus findings and complications after treatment were analyzed and compared between the CRVO-derived NVG and DR-derived NVG.All patients underwent panretinal photocoagulation,improving microcirculation therapy,anti-glaucoma (drug or surgery) and causative disease treatment,and some of them received vitrectomy or/and cataract surgery.Two eyes from each group received intravitreal injection of ranibizumab.The follow-up time in both groups was (14.00±10.13) months and (17.89±12.52) months,respectively.Results The median time of underlying disease was 3.3 months (2 weeks to 6 months) in the CRVO patients and 11.1 months (4 to 36 mouths) in the DR patients,with a significant difference between them (Z =-2.40,P＜0.05).CRVO-derived NVG progress was much faster than that of DR-derived NVG.The number of the eyes with visual acuity improvement after treatment was 2 in the CRVO-derived NVG and 15 in the DR-derived NVG;while the number of the eyes with unchanged or worse visual acuity was 8 and 4 in the CRVO-derived NVG eyes and the DR-derived NVG eyes (x2 =9.38,P＜0.01).The difference of IOP in pre-and post-treatment was (37.00±9.91)mmHg in the CRVOderived NVG eyes and (8.92±12.05)mmHg in the DR-derived NVG eyes,showing a significant difference between them (t =6.30,P＜0.01).In the CRVO-derived NVG eyes,optic disc edema,retinal hemorrhage,and vein dilatation were seen in 6 eyes,and mild optic disc edema and retinal hemorrhage were observed in 4 eyes.After treatment,fundus could not be seen in 4 eyes,in other 2 eyes optic disc and retinal laser spots were unclearly observed.In addition,pale optic disc and retinal vessel occlusion appeared in 2 eyes,and silver wire-like arteries exhibited in 2 eyes.In pre-treated DR-derived NVG eyes,fundus could not be seen in 8 eyes and Ⅲ-Ⅳv stages of DR findings appeared in 11 eyes.After treatment,retinopathy was stabilized in 16 eyes of 15 cases.Advanced retinopathy(V-Ⅵ stages of DR findings) was revealed in 3 eyes of 3 cases.The incidence of the complication after treatment was 100.0％ in the CRVO-derived NVG eyes and 21.1％ in the DR-derived NVG eyes (x2=5.18,P＜0.05).Conclusions The clinical characteristics of NVG secondary to CRVO and DR are variable,an appropriate treatment option should be selected according to different features of NVG.

Objective To investigate the infections caused by Staphylococcus aureus carrying Panton-Valentine leukocidin(PVL)genes.Methods 26 isolates of Staphylococcus aureus carrying Panton- Valentine leukocidin(PVL)genes were determined by multiplex PCR.Multilocus sequence typing(MLST) was used to determine the STs of the isolates.The genotypes of SCCmec were also determined by another multiplex PCR in the isolates of methicillin-resistant Staphylococcus aureus(MRSA).Results Among 26 isolates,there were 6 isolates of ST88 MRSA,7 isolates of ST88 methicillin-susceptible Staphylococcus aureus (MSSA),5 isolates of ST239 MRSA,5 isolates of ST398 MRSA,1 isolate of ST25 MRSA,1 isolate of ST30 MRSA and 1 isolate of ST59 MRSA.20 isolates were hospital-acquired(HA)which mainly caused pulmonary infection and post-operative pyogenic infection.6 isolates were community-acquired(CA)which mainly caused soft tissue necrosis.Among 19 isolates of MRSA,ST88-SCCmec Ⅲ A,ST239-SCCmec Ⅲ,ST398- SCCmec Ⅳ and ST398-SCCmec Ⅲ were main types.26 isolates were isolated from 14 wards.ST88-SCCmec Ⅲ A-MRSA caused clone spread in maternity department in our hospital.Conclusion ST88,ST239 and ST 398 are main STs in Staphylococcus aureus carrying PVL in our hospital.The isolates not only cause nosocomial infections but also cause community infection.

Objective To investigate the genetic polymorphism of mec Ⅰ in the clinical isolates of methicillin-resistant Staphylococcus anreus(MRSA).Methods 40 isolates(MRSA)carrying mecA gene were selected randomly from the clinical isolates of Staphylococcus anreus from Jan,2005 to Aug,2006 in our hospital.The mec Ⅰ gene was detected by PCR followed with sequencing.Staphylococcal cassette chromosome mec(SCCmec)in MRSA were detected by multiplex-PCR.Agar dilution method was used for determining the MICs of oxacillin against MRSA.Results 35 of 40(87.5%)MRSA carried mec Ⅰ gene.All isolates carrying mec Ⅰ gene have mecI 202C→T substitution,which resulted in Gln at 68 aminophenol position replaced by stop condon.32 isolates carried single point mutation.3 isolates carried double-point mutation,including additonal A at 3 positon,A→C at 41 position and C→T at 142 position beside C→T at 202 position,respectively.Among 35 isolates carrying mec Ⅰ gene,there were 27 isolates of SCCmec Ⅲ, 7 isolates of SCCmec Ⅲ A and 1 isolate of SCCmec Ⅱ.Among 5 isolates with deletion of mec Ⅰ gene,there were 3 isolates of SCCmecⅣ,1 isolate of SCCmec Ⅰ and 1 isolate of non-known SCCmec tpye.The MICs of oxacillin were 256-512 ?g/ml,≥512 ?g/ml and 8-256 ?g/ml in 31 isolates with single point mutation at 202 position in mec Ⅰ gene,3 isolates with double-point mutation in mecI gene and 5 isolates with deletion of mec Ⅰ gene,respectively.1 isolate with single point mutation in mec Ⅰ gene had contrary result(MIC

OBJECTIVETo study gene expression profiling in human type I and II endometrial carcinoma.

METHODSSix Affymetrix human genome genechips were utilized to investigate the differences in gene expression profiles between type I and II endometrial carcinoma with bioinformatic analysis.

RESULTSMany genes were highly expressed in estrogen-dependent endometrial carcinoma, and some of them were involved in the metabolism and conversion of estrogen, while some others in estrogen regulation. CYP2C9, for instance, was involved in the conversion of estrogen sulfate to 16-hydroxy sulfate metabolite, DDC in estrogen-dependent pathogenesis of endometrial carcinoma possibly by DDC interaction with AR to enhance steroid receptor transcription.

CONCLUSIONHigh expression of these genes in estrogen-dependent endometrial carcinoma may provide insights into their roles in the pathogenesis and prognosis of this malignancy.

Adenocarcinoma ; genetics ; pathology ; Adenocarcinoma, Clear Cell ; genetics ; pathology ; Endometrial Neoplasms ; classification ; genetics ; pathology ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Microarray Analysis ; Reverse Transcriptase Polymerase Chain Reaction

·AIM: To investigate the visual recovery factors in patients with complex traumatic and non-traumatic retinal detachment. ·METHODS: According to the history of ocular trauma before admission, 135 patients with complex retinal detachment were divided into traumatic group ( n=66, 66 eyes) and non-traumatic group (n=69, 69 eyes). The results of visual recovery and complications were compared between the two groups. Logistic regression was used to analyze the related factors of visual recovery in the two groups. ·RESULTS: There was no significant difference in the recovery rate between the two groups within 24h after surgery (77. 3% vs 78. 3%, P>0. 05), and there was no significant difference in the total incidence of complications at 3mo after surgery(18. 2% vs 17. 4%, P>0. 05). Multifactorial Logistic regression analysis showed significant correlation between age, injury type, time since retinal detachment, rage of retinal detachment, preoperative vitreous blood,proliferative vitreoretinopathy ( PVR ) and postoperative visual recovery in patients with complex traumatic retinal detachment ( P < 0. 05 ); age, time since retinal detachment, rage of retinal detachment and macular status were significantly associated with visual recovery in patients with complex non-traumatic retinal detachment (P<0. 05). ·CONCLUSION: Age, time since retinal detachment and rage of retinal detachment were significantly associated with traumatic and non-traumatic retinal detachment. The injury type, preoperative vitreous hemorrhage, PVR were significantly correlated with the visual recovery of traumatic retinal detachment patients. The condition of macular was significantly associated with the visual recovery of non-traumatic retinal detachment patients.

Objective To explore the surgical skills and efficacy of microsurgical excision of large acoustic neurinoma via the suboccipital retrosigmoid keyhole approach. Methods Fifty-nine patients with large acoustic neurinoma (≥3 cm) underwent microsurgical resection via suboccipital retrosigrmoid keyhole approach. The intraoperative position of the operative bed and angle of the microscope were adjusted to expose the tumors sufficiently. The duramater was sutured tightly and the bone flap was replaced and fixed. Results Of the 59 patients, 53 (89.8%) received a total resection of the tumors, and 6 (10.2%) subtotal resection. No patient died. The facial nerve was anatomically preserved in all the patients. Forty-five patients (76.3%) suffered from mild or moderate facial palsy after the surgery, and the symptoms were improved significantly after expectant treatment. Fifteen patients preserved partial hearing. No subcutaneous water accumulation or cerebrospinal fluid leakage occurred.Conclusion Microsurgery via suboccipital retrosigmoid keyhole approach is a favorable treatment for large acoustic neurinomas with low morbidity and mortality,which can effectively protect the function of the acoustic and facial nerves.

OBJECTIVETo investigate the effects of arginine enriched enteral nutrition (EN) on nutritional status and cellular immunity of severely burned patients.

METHODSRandomized, single blind, parallel and positive control investigation was employed in the study. Thirty severely burned patients were divided into enteral immune nutrition (EIN) group and EN group. Sixteen patients in EIN group received enteral nutrition enriched with arginine, while the other 14 patients in EN group received standard enteral nutrition. Nutritional support was continued for 14 days. Gastrointestinal reaction of patients in 2 groups was observed. Fasting venous blood was drawn from patients of both groups before receiving nutrition treatment and on the morning of 7th, 14th day of treatment. Level of serum protein, hepatic function parameters, renal function parameters, fasting-blood glucose, and subpopulations of T lymphocytes in peripheral blood were determined.

RESULTS(1) Incidence of gastrointestinal side effect in EIN group (25.0%) was close to that of EN group (21.4% , P > 0.05). (2) Compared with pre-treatment days, levels of prealbumin and transferrin in serum of patients in 2 groups on 7th and 14th post-treatment days were significantly increased (P < 0.05 or P < 0.01), but there was no significant difference between 2 groups. The level of total serum protein on 14th day of treatment of patients was significantly increased in both groups, and that of EIN group (66 +/- 7 g/L) was significantly higher compared with that in EN group (64 +/- 11 g/L, P < 0.05). The level of serum albumin (29 +/- 5, 32 +/- 5 g/L, respectively) of patients in EIN group on 7th and 14th day of treatment were significantly higher than that (26 +/- 4 g/L, P < 0.05) in pre-treatment days, however there was no significant difference in EN group. (3) There was no significant difference in respect of hepatic function, renal function, and fasting-blood glucose between pre-treatment and post-treatment periods in both groups (P > 0.05). (4) The ratio of CD4(+), CD8(+) on 14th day of treatment in EIN group was close to that of pretreatment level. In EN group, cell percentage of CD4(+) significantly decreased, while that of CD8(+) significantly increased (P < 0.05), and CD4(+) was significantly higher [(56 +/- 8)%] in EIN group than that in EN group [(55 +/- 12)%, P < 0.05]. In both groups, cell percentage of CD3(+) was significantly higher than that in pre-treatment days (P < 0.05), while there was no obvious change in CD4(+)/CD8(+).

CONCLUSIONSArginine enriched enteral nutrition can effectively improve nutritional status and cellular immune function of burn patients.

Adolescent ; Adult ; Aged ; Arginine ; administration & dosage ; Burns ; immunology ; therapy ; Enteral Nutrition ; methods ; Female ; Humans ; Immunity, Cellular ; immunology ; Male ; Middle Aged ; Nutritional Status ; Single-Blind Method ; Treatment Outcome ; Young Adult