Objective To observe the morphologic protection effect of intra-articular injection of osteoprotegerin (OPG)on articular cartilage in a rabbit model of osteoarthritis (OA).Methods Sixty male New Zealand rabbits were randomly divided into 3 groups:OPG group (n=20),sham-operated group (n=20) and PBS group (n=20).In OPG group and PBS group,each rabbit underwent anterior cruciate ligament transection (ACLT) in left knee joint,then 0.1 ml OPG solution or PBS were injected into the left knee for 8 weeks (5 times a week) in OPG group and PBS group,respectively.In sham-operated group,the anterior cruciate ligament was just exposed without transection,and then the incision was closed.All rabbits were sacrificed 12 weeks after operation,and the left knee joints were obtained.The Pelletier score and Mankin score were used to evaluate the macroscopic and microscopic cartilage morphology.Results The score of femoral condyle cartilage and tibial plateau cartilage was 1.80±0.89 and 1.80±0.77 in OPG group,respectively,and 3.10±0.97 and 3.20±0.77 in PBS group.However,there was no statistical difference in Pelletier score between the sham-operated group and the OPG group.Consistent with the macroscopic data,the OPG group has a significantly lower Mankin score involving cartilage structure (2.65±0.88),chondrocyte (1.35±0.71),Safranin O staining (1.83±0.83),tidemark integrity (0.30±0.47) and total score (6.13±1.97) compared with the PBS group (4.52±1.09,1.85±0.63,2.80±0.75,0.65±0.49,and 9.83±1.98,respectively).The OPG group has a significantly higher total Mankin score compared with the sham-operated group (4.80±1.25),however,there were no statistical differences in four subitem scores between the two groups.Moreover,the cartilage thickness in OPG group was 371.84±38.94 μm,255.09±74.82 μm in PBS group,and 404.68±15.97 μm in the sham-operated group,and the differences were statistically significant between three groups.Conclusion OPG has a protective effect on articular cartilage and can slow the progression of OA by reducing the grade of synovitis,decreasing the volume of osteophytes,and suppressing the loss of cartilage thickness.

Objective To observe the effect of intra-articular injection of berberine on the expression of matrix metalloproteinase (MMP)-13,ADAMTS-5,Aggrecan and Collagen Ⅱ in the cartilage of rabbits with osteoarthritis.Methods Thirty male New Zealand White rabbits underwent bilateral anterior cruciate ligament transection (ACLT).On the basis of randomization one knee of each rabbit was treated with 0.3 ml 100 μmol/L berberine resolved in the normal saline (NS) (the experimental group) and the other knee was treated under the same schedule using NS (the placebo group) 4 weeks after transection,once a week for five weeks.Nine weeks after ACLT,all rabbits were killed and the knee joints were evaluated by histology and biochemistry.The mRNA expression of MMP-13,ADAMTS-5,Aggrecan and Collagen Ⅱ in the cartilage was analyzed using reverse transcription polymerase chain reaction (RT-PCR).All data were analyzed using Student's t test.Results Mankin histological evaluation showed that the extent and grade of cartilage damage in the experimental group (6.9±1.9) were less severe than the placebo group (9.3±1.2)(t=3.394,P＜0.01).The mRNA expression of MMP-13 and ADAMTS-5 decreased (160±54;166±47) (t=3.311,P＜0.01;t=2.651,P＜0.05) while that of Aggrecan (261±50) and Collagen Ⅱ (335±64) increased in cartilage compared to the placebo group (233±45;234±67;186±64;254±69),the differences were significant (t=2.941,P＜0.01;t=2.743,P＜0.05).The content of hydroxyproline (Hyp) and glycosaminoglycan (GAG) in cartilage [(23.5±2.8) μg/mg;(30±3) μg/mg] increased significantly in the experimental group (t=2.941,P＜0.01;t=2.743,P＜0.05) compared to the placebo group [(19.9±2.8) μg/mg;(27.4±2.9) μg/mg].Conclusion Berberine protects against cartilage degradation and inhibits the progression of osteoarthritis by suppressing MMP-13 and ADAMTS-5 expression.

Objective To study the effect of intra-articular injection of dehydroepiandrosterone (DHEA) on the balance of ADAMTS/TIMP-3 system in a rabbit osteoarthritis models.Methods Sixty rabbits were underwent bilateral anterior cruciate ligament transection (ACLT).Rabbits were randomizedn to the following treatment:one knee of each rabbit was treated with 100 μmol/L DHEA dissolved in dimethylsulphoxide (the experimental group) and the other knee was treated under the same schedule using dimethylsulphoxide (the control group) 4 weeks after transection,once a week for eight weeks.Twelve weeks after ACLT,all rabbits were killed after X-ray assessment and the knee joints were evaluated by gross morphology and histology.The concentration of hydroxyproline and glycosaminoglycan in the cartilage were analyzed.The mRNA expression of ADAMTS-4,ADAMTS-5,tissue inhibitor of metalloproteinases-3 (TIMP-3),transforming growth factor (TGF)-β1.Aggrecan and Collagen Ⅱ in the cartilage were analyzed using reverse transcription polymerase chain reaction (RT-PCR).The protein expression of aggrecan ARGxx and Collagen Ⅱ in the cartilage were analyzed by Western blot.Results By Mann-Whitney test,Gross morphologic scores on femoral condyle and tibial plateau in the control group were significantly higher than the experi-mental group (Z=-3.517,P＜0.01 ; Z=-2.518,P＜0.05).By unpaired Student's t test,histological evaluation showed that the grade of cartilage damage in the experimental group [(5.3±1.2) μg/ml] were less severe than that in the control group (10.1 ± 1.3,P＜0.01).The concentration of hydroxyproline [(5.7±0.3,23.6± 1.7) μg/ml] and glycosaminoglycan (30±4) in the experimental group increased significantly when compared with the control group [(4.6±0.5),(18.5±1.4),(24±4) μg/ml,P＜0.01].The mRNA expression of ADAMTS-4 (0.15±0.03)and ADAMTS-5 (0.10±0.04) in the experimental group decreased significantly compared with the control group (0.29±0.08,0.15±0.05; all P＜0.05).The mRNA expression of TIMP-3 (0.85±0.10),TGF-β1(1.2±0.4),Aggrecan (0.87±0.31) and Collagen Ⅱ (2.74±0.59) in the experimental group increased significantly when compared with the control group (0.70±0.13,0.8±0.4,0.49±0.16,2.2±0.5; all P＜0.05).The protein expression of Aggrecan ARGxx (0.53±0.10) in the experimental group decreased significantly when compared with the control group (0.81±0.12,P＜0.01).The protein expression of Collagen Ⅱ (2.3±0.7) in the experimental group increased significantly when compared with the control group (1.7±0.5,P＜0.05).Conclusion DHEA protects against cartilage degradation and inhibits the progression of OA,TGF-β1,Aggrecan and Collagen Ⅱ in cartilage may be the mechanism of the protective effect of DHEA on OA.

Objective To eveluate the clinical curative effects of combination of Ma Yinglong She Xiang Zhi Chuang Gao,Jin Xuan Zhi Ke Xun Xi Xan,Diosmin,Macrogol 4000 powder in treating postoperative complications of ring mixed hemorrhoids.Methods Ninty cases of postoperative ring mixed hemorrhoids patients were divided into two groups randomly from January 2008 to June 2009.Experimental group:From the first day on Jin Xuan Zhi Ke Xun Xi San 55 g and the 1000 mL boiling water were added flushing,and Maying Long She Xiang Zhi Chuang Gao 2.5 g,2 times daily.The Macrogol 4000 powder 10 g and water 200 mL were admistrated orally,Diosmin 1.0 g orally,2 times daily.Oral administration of two kinds of medications was done each two hours.Control group:Using 1:100 Sterile warm salt water hip bath,and Ma Yinglong she xiang zhi chuang gao 2.5 g,2 times daily;Phenolphthalein tablets 100 mg orally,2 times daily.Results The experimental group surpassed the control group in the anus ache,the hemorrhage,edema (P<0.05).The heal time reduced obviously(P<0.01).Conclusion To combination of Ma Yinglong She Xiang Zhi Chuang Gao,Jin Xuan Zhi Ke Xun Xi San,Diosmin,Macrogol 4000 powder has the distinct improvement in the anus ache,the hemorrhage,dropsy of ring mixed hemorrhoids and reduces the injured area heal time obviously.

Objective To investigate the relationship between baseline serum uric acid and the severity of coronary artery disease ( CAD ) in the first-degree relatives or non-first-degree relatives of men with type 2 diabetes. Methods Three hundred and eighty-one men with negative coronary angiography for the first time were divided into diabetes and non-diabetes groups and followed-up for 5 years. The primary outcome was acute coronary syndrome suspected during subsequent 5 years, and the coronary angiography was conducted simultaneously. The severity of CAD was assessed by the coronary stenosis index ( CSI) and the number of coronary lesion vessels. Results In normal blood glucose group, serum uric acid was higher in the first-degree relatives of diabetics compared with non-first-degree relatives(P<0. 01), along with higher morbidity of CAD, CSI, and coronary lesion vessels (all P<0.01). Correlation analysis showed that CSI(r=0. 250, P=0. 041) and coronary lesion vessels(r=0. 252, P=0. 040) in non-diabetics group were associated with baseline levels of serum uric acid. Conclusion The elevation of serum uric acid was closely related to subsequent CAD, especially in first-degree relatives of male with type 2 diabetes, which could be used as an early indicator for CAD prediction.

Objective To investigate the impact of enhanced recovery after surgery (ERAS)on immune function in patients undergoing laparoscopic hepatectomy.Methods Sixty patients undergoing laparoscopic hepatectomy (34 males,26 females,aged 38-57 years,ASA Ⅰ or Ⅱ),were randomly divided into two groups:enhanced recovery after surgery group (group E)and non-enhanced recovery after surgery group (group C).The patients in group E received enhanced recovery after surgery,while the patients in group C received routine perioperative management and anesthesia methods.The operation method and time,the volume of bleeding,the intraoperative fentanyl con-sumption,the volume of fluid input,the preoperative and postoperative CVP and temperature were recorded in the two groups.Blood samples were obtained before induction (T0 ),at the end of opera-tion (T1 ),on day 1 (T2 ),day 3 (T3 ),day 7 (T4 )after operation for determination of plasma con-centration of IgA,IgM,IgG and the percentages of T lymphocyte subsets (CD3+ ,CD4+ ,CD8+ ) and CD4+/CD8+ ratio were detected with flow cytometry.Furthermore,the visual analogue scale (VAS)score and Ramsay score were evaluated 4 hours,8 hours,24 hours and 48 hours after opera-tion in two groups.Results Compared with group C,the intraoperative fentanyl consumption,the volume of fluid input and the postoperative CVP in group E were significantly decreased,while the postoperative temperature was significantly increased (P < 0.05 ).Compared with T0 , the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ ratio and the plasma concentration of IgA,IgM,IgG in group E on T1-T3 were significantly decreased,the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ratio and the plasma concentration of IgA,IgM,IgG in group C on T1-T4 were significantly decreased (P <0.05).Compared with group C,the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ ratio and the plasma concentration of IgA,IgM,IgG in group E on T1-T4 were significantly increased (P <0.05),the visual analogue scale (VAS)score 4 hours,8 hours,24 hours after operation were signifi-cantly decreased (P <0.05).The comparision of Ramsay scores at all the time points between two groups were similar.Conclusion ERAS applied to patients undergoing laparoscopic hepatectomy can reduce the intraoperative fentanyl consumption,prevent the occurrence of hypothermia and provide satisfactory postoperative analgesia,which can significantly improve the immune function in patients.

Objective To evaluate the effect of stellate ganglion block (SGB) on cellular immune function in diabetic rats.Methods Healthy male Sprague-Dawley rats,aged 3 months,weighing 240-280 g,were used in this study.Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.Forty-eight rats with diabetes mellitus were randomly divided into 2 groups (n=24 each) using a random number table:diabetes mellitus group (group DM) and group SGB.Another 24 healthy rats,aged 3 months,were selected and served as control group (group C).At 1 week after successful establishment of the model,unilateral transection of cervical sympathetic trunk (TCST) was performed in group SGB,while the right cervical sympathetic trunk was only exposed in C and DM groups.Before TCST (T0) and on 1,3,7 days after TCST (T1-3),6 rats were randomly selected from each group,and blood samples were collected from the inferior vena cava for determination of the blood glucose,plasma norepinephrine (NE) concentrations (by enzyme-linked immunosorbent assay),and levels of T lymphocyte subsets CD3+,CD4+ and CD8+ in whole blood (using FACSCalibur flow cytometer).C D4+/CD8+ratio was calculated.The rats were weighed before sacrifice,and the rats were sacrificed to obtain the thymus which was weighed.The thymus index (thymus weight/body weight) was calculated.Results Compared with group C,the blood glucose was significantly increased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly decreased at T0-3 (P＜0.05),and no significant change was found in CD8+ levels in DM and SGB groups (P＞0.05),the plasma NE concentrations were significantly decreased at T1-3 in group SGB (P＜0.05),and no significant change was found in plasma NE concentrations in group DM (P＞0.05).Compared with group DM,the blood glucose and plasma NE concentrations were significantly decreased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly increased at T1-3 (P＜0.05),and no significant change was found in CD8+ levels in group SGB (P＞0.05).Conclusion SGB can improve the cellular immune function in diabetic rats.

Objective:To explore the cleansing effect of Nitric Oxide (NO) sustained-release silica nanoparticles (short for NO sustained-release agent) on endoscopic biofilm and its clinical application.Methods:A total of 160 clinical endoscopes were randomly divided into two groups: the cleansing agent group (80 pieces, disinfected with cleansing agents), NO group (80 pieces, disinfected with NO sustained-release agent). A biofilm model of Pseudomonas aeruginosa was constructed and used as the control for phosphate buffered solution (PBS) treatment. A biofilm model of Pseudomonas aeruginosa on the surface of endoscopic lumen was built first in vitro. Scanning electron microscopy was then used to observe the microstructure of biofilm after treatment with NO sustained-release agent. Viable counting method was used to evaluate the cleansing effect of NO sustained-release agent on biofilm. Finally, at the clinical level, the actual disinfection effect of NO sustained-release agent on clinical endoscopy was evaluated by detecting the protein residues, viable counting and adenosine triphosphate (ATP) biofluorescence detection. Results:The scanning electron microscopy showed that the biofilm was intact in the model group, but scattered bacteria were observed on the biofilm surface in the NO group and the detergent group. Compared with the model group [(4.86±2.67)×10 6(colony-forming units, CFU)/mL], the standard CFUs of the NO group [(1.37±0.61)×10 4CFU/mL] and the detergent group [(1.31±0.21)×10 5CFU/mL] were significantly lower (detergent group VS model group, P=0.009; NO group VS model group, P=0.008), and there was significant difference between the detergent group and the model group ( t=9.53, P=0.000 6). The levels of residual proteins in the endoscopic lumens before and after the treatment were 8.03±1.47 mg/mL and 0.50±0.37 mg/mL in the NO group, 8.01±1.51 mg/mL and 0.91±0.52 mg/mL in the detergent group with significant difference ( P<0.01), and the reduction effect of the NO group was more significant. The disinfection of NO group and cleaning agent group was within the qualifying range, but the ATP bioluminescence value, protein residue and colony count of NO group (78.56±42.59 RLU, 0.50±0.37 mg/mL, 7.55±4.56 CFU) were significantly lower than those of detergent agent group (120.80±54.00 RLU,0.91±0.52 mg/mL,11.50±4.75 CFU, P<0.01). Conclusion:NO sustained-release agent can effectively clear endoscopic biofilm and further improve the disinfection effect on endoscopes, which may be of great significance for improving the effects on treatment and prognosis of patients.

OBJECTIVETo determine the function of twin-arginine translocation system (Tat) and gene cluster in Vibrio strains and to analyze the homology of tat gene cluster among different Vibrio spp. strains based on N16961 and tatABC mutant strains N169-dtat.

METHODSDifferent serotypes of biotype strains of Vibrio spp. were selected to detect the transcription of 4 genes of Tat transport system and upstream ubi aarF gene and downstream cyt551 gene by the total RNA reverse transcription and homologicity of the gene cluster by sequencing analysis.

RESULTSOur results showed that the 4 genes of tat cluster (tatA, tatB, tatC, and tatE) were intragenic and co-transcribed. We found that ubi aarF gene could be co-transcribed with tatA, tatB, but not with tatC. The electron transport chain and energy metabolism-related genes, cytochrome C551 peroxidase gene, and 4 genes located at upstream of tatABC operon were not transcribed with tatABC. Although the co-transcription between ubi aarF and tatAB was blocked in N169-dtat strain, they were still transcribed separately. Homologous analysis of genes of tat cluster in different types of Vibrio cholerae showed that tat gene cluster was a very conservative.

CONCLUSIONThe ubi and aarF gene might be co-transcribed with genes of tat cluster in Vibrio cholerae, which and the close relationship showed that they might play a key function in Vibrio cholerae.

Arginine ; Bacterial Proteins ; Cytochrome c Group ; Membrane Transport Proteins ; Vibrio cholerae

OBJECTIVETo analyze virulence genes and molecular characteristics of Vibrio parahaemolyticus isolated from sporadic cases with diarrhea in tow sentinel hospitals of Shanghai, 2010-2012.

METHODSA total of 2 729 stool samples were collected from two surveillance sentinel hospitals in Shanghai 2010-2012. Vibrio parahaemolyticus strains were isolated and identified from diarrhea out patients using TCBS agar plates and biochemical reactions. Thermostable direct hemolysingene (tdh), thermostable-related hemolysin gene (trh), hemolysin gene (tlh) were detected by multiplex PCR method. Isolates were analyzed by PFGE and MLST. The PFGE profiles were analyzed using BioNumerics software.

RESULTSA total of 30 clinical Vibrio parahaemolyticus strains isolated from 2 729 stool samples. The anually Vibrio parahaemolyticus isolation rate during 2010 to 2012 were 1.1%(11/973), 1.0%(11/1 120) and 1.3%(8/636) respectively. The PCR positive rates of virulence genes tlh, tdh and trh were 100%, 97% and 0 respectively. The Vibrio parahaemolyticus strains were divided into 13 PFGE types (P1-P13)and 3 ST types (ST-189, ST-799, ST-3). Among 13 PFGE types, P4 was the main PFGE type, accounting for 30%(9/30). P9, P10 were accounting for 12% (4/30) respectively, P1, P2, P12, P13 were accounting for 7%(2/30) respectively, the others types were 3%(1/30) respectively. MLST analysis results showed there are three ST types, ST3 was 84%(25/30), ST189 and ST799 were accounting for 13% (4/30) and 3% (1/30) respectively.

CONCLUSIONThe infection rate of Vibrio parahaemolyticus was not very high from 2010-2012 in Shanghai, all strains were positive for tlh and negative for trh. ST3 was the major type of Vibrio parahaemolyticus.

China ; Diarrhea ; Genotype ; Hemolysin Proteins ; Hospitals ; Humans ; Multilocus Sequence Typing ; Outpatients ; Polymerase Chain Reaction ; Sentinel Surveillance ; Vibrio Infections ; Vibrio parahaemolyticus ; Virulence