OBJECTIVETo compare the clinical effects between hip anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery and posterior K-L approach combined with quadratus femoris bone flap transplantation for the treatment of femoral neck fracture of Garden III-IV in young and middle-aged patients.

METHODSFrom January 2004 to January 2011,46 patients with femoral neck fractures were treated by two kinds of operation. Among them, 20 cases were treated with anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery, included 12 males and 8 females with an average age of (32.1 ± 7.3) years old, involved 12 cases of Garden III and 8 cases of Garden IV. The other 26 cases were treated with posterior K-L approach combined with quadratus femoris bone flap transplantation, included 20 males and 6 females with an average age of (37.8 ± 6.9) years old, involved 16 cases of Garden III and 10 cases of Garden IV. The index of hospitalization (hospitalization time, total cost, operative time, intraoperative blood loss, postoperative complications), the quality index of operation (fracture reduction, position of internal fixation, fracture healing time, nonunion and femoral head necrosis) of two groups were observed and compared. Hip joint function were evaluated by Harris score.

RESULTSAll patients were followed up from 28 to 41 months with an average of 36 months. The intraoperative blood loss of group S-P (92.3 ± 10.4) ml was less than that of group K-L (132.4 ± 11.2) ml, there was significant difference between two groups (P < 0.05). The operation time of group S-P (81.4 ± 9.2) min was more than that of group K-L (67.1 ± 4.5) min, the difference was statistically significant (P < 0.05). One case in group S-P and 9 cases in group K-L appeared postoperative complications, there was significant difference between two groups (P < 0.05). The fracture healing time of S-P group (83.5 ± 7.3) d was shorter than that of group K-L (103.2 ± 12.6) d, there was significant difference between two groups (P < 0.05). At 30 months after operation, there were significant difference in Harris scoring between two groups (P < 0.05).

CONCLUSIONAnterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery for treatment of femoral neck fracture of Garden III-IV of young and middle-aged patients, it has characteristics in clear anatomic and easy to operate. As compared with K-L approach, S-P approach can better reserve residual blood supply of femoral neck. While combining with the iliac bone flap transplantation with deep circumflex iliac artery, it could better reconstruct the blood supply of femoral neck to promote fracture healing.

Adult ; Case-Control Studies ; Female ; Femoral Neck Fractures ; physiopathology ; surgery ; Fracture Healing ; Humans ; Iliac Artery ; Male ; Middle Aged ; Surgical Flaps ; transplantation

OBJECTIVETo set up a venous congested flap model to study the mechanism of necrosis through long-term microcirculation observation.

METHODSA specially deviced chamber was assembled to one side of the ears in an adult white rabbit, about 7 approximately 10 days after the operation the congested flap model was made and the microcirculatory status of the flap was dynamically observed under a vivo-microscope for a long time.

RESULTSThe venous crisis phenomenon of flap was well studied and the microcirculation of the flap was observed carefully, finally the variational rule of the congestion flap microcirculation was made clear.

CONCLUSIONSThe model could well simulate the venous crisis flap in clinic, and the microcirculation could also be observed for a long time.

Animals ; Disease Models, Animal ; Female ; Male ; Microcirculation ; Rabbits ; Surgical Flaps ; blood supply ; pathology ; Veins ; pathology

cuses tumors. Survivin seems to be a promising marker for analyzing clinical stages and predicting the prognosis of TCC.

Objective To explore the adeno-associated virus (AAV)gene transduction and cellular endocytosis mediated by ultrasound combined with microbubbles in two types of cells.Methods HeLa and NIH/3T3 cells were infected by rAAV2-EGFP at a concentration gradient to get the optimal concentrations for enhancement.At these concentrations,HeLa and NIH/3T3 cells were infected by rAAV2-EGFP mediated by ultrasound combined with microbubbles.The gene transduction efficiency were observed and measured by fluorescence microscopy and flow cytometry at 48 h after treatment.The cell viability was tested by CCK-8.The number and distribution of cellular clathrin-coated endocytic pits were observed by confocal fluorescence microscopy and transmission electron microscopy on 45 min after treatment.Results The optimal concentrations for HeLa and NIH/3T3 cells were 2000 v.g./cell and 10000 v.g./cell.Ultrasound combined with microbubbles significantly enhanced the transduction efficiency of rAAV2-EGFP (P <0.01) without significant cell viability decrease (P > 0.05 ).Confocal fluorescence microscopy and transmission electron microscopy demonstrated that clathrin-coated endocytic pits were more obviously increased in ultrasound combined with microbubbles mediated AAV transduction group than AAV transduction group. Conclusions Ultrasound combined with microbubbles can efficiently enhance the gene transduction of AAV,whose cellular transportation depends on cellular endocytosis,in two types of cells.Stimulating cellular endocytosis might be one of the mechanisms of enhanced cellular transportation of AAV mediated by ultrasound combined with microbubbles.

Objective To compare the levels of the serum pepsinogen (PG) in the gastric diseases, and explore the diagnostic value in gastric diseases. Methods Two hundred and fourteen patients who had undergone endoscopy were selected, and the patients were divided into 3 groups according to the results of endoscope pathological diagnosis:chronic superficial gastritis (CSG) group ( 70 cases), chronic atrophic gastritis (CAG) group (86 cases) and gastric cancer (GC) group (58 cases). The quantitative chemiluminescence method was used to test serum PGⅠand PGⅡ, and the PGⅠ/PGⅡratio (PGR) was calculated. Results The PGⅠin GC group was significantly higher than that in CAG group: (78.41 ± 55.42) μg/L vs. (53.10 ± 30.08) μg/L, and there was statistical difference (P<0.05). There was no statistical difference in PGⅠbetween GC group and CSG group (P>0.05). The PGⅡin GC group was significantly higher than that in CAG group and CSG group: (23.26 ± 17.80) μg/L vs. (13.12 ± 10.23) and (13.78 ± 9.26) μg/L, the PGR was significantly lower than that in CAG group and CSG group:3.67±2.03 vs. 4.88 ± 1.82 and 5.24 ± 1.88, and there were statistical differences (P<0.05). Helicobacter pylori (Hp) was detected in 165 patients, with positive in 29 cases (Hp positive group) and negative in 136 cases (Hp negative group). There was no statistical difference in PG Ⅰ between Hp negative group an Hp positive group:(60.46 ± 45.49)μg/L vs. (72.41 ± 31.85)μg/L, P>0.05. The PGⅡin Hp positive group was significantly higher than that in Hp negative group: (19.58 ± 1.57) μg/L vs. (14.09 ± 13.21) μg/L, the PGR was significantly lower than that in Hp negative group: 3.82 ± 0.18 vs. 4.99 ± 0.18, and there were statistical differences (P<0.05). Conclusions Compared with that in the CSG and CAG patients, the PG Ⅱ in GC patients increases significantly, while PGR descends significantly, but PG Ⅰ has no correlation with the risk of GC. The PG Ⅱ combined with PGR can predict people with high risk of GC, and help with the judgment of Hp infection.

[ABSTRACT]AIM:TostudytheeffectofsmallinterferingRNA(siRNA)ontheexpressionofbeta2-microglo-bulin (β2M) in pre-differentiated bone marrow mesenchymal stem cells (BMSCs).METHODS: The β2M siRNA was transfected into the pre-differentiated BMSCs with Lipofectamine 2000.BMSCs were divided into transfection group , blank control group and negative control group .The expression of β2 M at mRNA and protein levels was determined by real-time qPCR, Western blotting and laser confocal microscopy .The productions of aggrecan and type II collagen in pre-differentia-ted BMSCs were determined by toluidine blue staining and type Ⅱcollagen immunofluorescence .RESULTS:The results of real-time qPCR, Western blotting and laser confocal microscopy showed that siRNA successfully inhibited the expression ofβ2 M at mRNA and protein levels in the pre-differentiated BMSCs .The results of toluidine blue and type Ⅱcollagen im-munofluorescence staining showed that siRNA does not affect the productions of aggrecan and type Ⅱ collagen in the pre-differentiated BMSCs .CONCLUSION:siRNA targeting β2 M reduces the expression of β2 M in the pre-differentiated BM-SCs and does not affect the chondrocyte characteristics of pre -differentiated BMSCs .

[ABSTRACT]AIM:Toinvestigatetheinhibitoryeffectsofresveratrolonchondrosarcomaandtherelationwith mitochondrial and PI3K/Akt pathways.METHODS:Chondrosarcoma SW1353 cells were treated with resveratrol at con-centrations of 25, 50 and 100 μmol/L for the time intervals of 24 h, 48 h and 72 h.The viability and apoptosis of the SW1353 cells in the presence or absence of resveratrol were analyzed by CCK 8 assay and Hoechst 33258 staining , respec-tively.The protein levels of Bcl-2, Bax, activated caspase-3, Akt and p-Akt were detected by Western blotting .The cell migration ability was determined by wound scratch assay .RESULTS:Exposure of the cells to resveratrol resulted in a de-crease in the cell viability in a dose-and time-dependent manner (P<0.05).visible nuclei with apoptotic characteristics in resveratrol group were observed .The protein levels of activated caspase-3 and Bax were increased , and Bcl-2 and p-Akt were decreased compared with control group .The total Akt were not significantly changed .Resveratrol also significantly re-duced the migration of tumor cells .CONCLUSION:Resveratrol induces apoptosis of chondrosarcoma , which plays a role of part through mitochondrial and PI 3K/Akt signaling pathways .

BACKGROUND:Many in vivo and in vitro experiments indicate that hypoxic co-cultures promote stem cells differentiate into chondrocytes.
OBJECTIVE:To evaluate the influence of hypoxia on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes.
METHODS:Adipose-derived stem cells and articular chondrocytes were mixed at the ratio of 3:1, then the mixed cells were seeded onto poly(lactic-co-glycolic acid)-gelatin scaffold at the ultimate concentration of 5.0×1010/L. The cells were cultured in normoxia (20%O 2 ) and hypoxic (5%O 2 ) conditions for 6 weeks. After culture, hematoxylin and eosin staining was performed for histological structure analysis, and alcian blue staining was used to evaluate glycosaminoglycan synthesis. Type II col agen expression was detected by immunohistochemistry staining. The content of DNA, glycosaminoglycan and hydroxyproline in the scaffold-cellcomplex was measured.
RESULTS AND CONCLUSION:In the hypoxia group, hematoxylin-eosin staining showed the formation of massive cells and extracellular matrix;alcian blue staining showed massive glycosaminoglycan formation;immunohistochemistry staining detected strongly positive expression of col agen type II, the content of DNA, glycosaminoglycan and hydroxyproline was higher than the normoxia group. Hypoxia promotes in vitro chondrogenic differentiation of co-cultured adipose-derived stem cells and articular chondrocytes. .

Objective To investigate the expression level of adenosine 2A receptors in peripheral blood lymphocytes and the correlation with disease progression of Parkinson's disease (PD).Methods In this retrospcctive study,out patients with PD(42 cases)and healthy controls(32 cases) were recruited at Beijing Hospital.The expression level of A2A receptors in peripheral blood lymphocytes was detected by flow cytometry.The concentration of free A2A receptors in plasma was detected by enzyme linked immunosorbent assay (ELISA).The expression of A2A receptors and plasma free A2A receptors in peripheral blood lymphocytes of the PD group and the control group was compared.Multivariate regression analysis and single factor correlation analysis were performed on sex,age,course of disease,duration of medication,drug type and dose,motor complications,and PD unified Parkinson's disease rating scale (UPDRS)score.Results A2A receptor expression in lymphocytes was significantly higher in the PD group than in the control group (8.96 ± 4.73)％ vs.(5.39±2.42)％ (t=4.210,P＜0.05).There was no significant difference in A2A receptor concentrations in plasma between the two groups (1.82 ± 1.91) μg/L vs.(1.15 ± 0.71) μg/L(t=1.078,P＞0.05).In the PD group,A2A receptor expression in lymphocytes in patients with motor complications was statistically lower than in patients without them (P＜ 0.05).Lymphocyte A2A receptor levels in the 5-9 years duration subgroup were significantly lower than those in the ＜5 years duration subgroup (Z=2.780,P＜0.01) and the≥10 years duration subgroup (Z=-2.149,P＜0.05).Conclusions The expression of A2A receptors in peripheral blood lymphocytes is correlated with PD.The expression of A2 A receptors in peripheral blood lymphocytes of patients with PD fluctuates with the occurrence of motor complications and the progression of disease.Further research is needed to establish A2A receptors as a biomarker for monitoring disease progression.

Objective: To analyse the expression of differential mRNA in the plasma exosomes in patients with latent tuberculosis infection (LTBI) and active tuberculosis (ATB) using high-throughput sequencing, so as to provide insights into differential diagnosis of LTBI and ATB. Methods: The plasma samples were collected from the patients treated at The Affiliated Hospital of Hangzhou Normal University, including 16 cases of LTBI and 21 cases of ATB. The exosomes were extracted by Invitrogen extracellular extracts purification kit, and the size and morphology of exosomes were observed by transmission electron microscope (TEM). The exosomes were identified by Western blotting. Total RNA was extracted from plasma exosomes using high-throughput sequencing, differential expression mRNA was identified, and gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed. Two differential mRNAs with the highest differential expression fold were selected, and five patients with ATB and three patients with LTBI were recruited for verification using real-time quantitative PCR. Results: The sequencing results of plasma exosomes showed that compared with ATB patients, 2 875 differentially expressed mRNAs were detected in exosomes of LTBI patients, of which 1 002 mRNAs were up-regulated and 1 873 mRNAs were down-regulated. The most significant differentially expressed downregulated and upregulated mRNA were M6PR and RGPD5, respectively. GO analysis and KEGG pathway analysis showed that differential mRNAs were enriched in protein serine kinase activity, rRNA binding molecular function, human cytomegalovirus infection, pancreatic cancer, endometrial cancer, insulin signaling pathway and FoxO signaling pathway. The real-time quantitative PCR showed that the expression of differential mRNA was consistent with sequencing. Compared with ATB patients, the relative expression level of M6PR in plasma exosomes in LTBI patients (0.954±0.212) was downregulated compared with that of ATB patients (2.168±0.226), while the relative expression level of RGPD5 (2.126±0.200) was upregulated compared with that of ATB patients (0.588±0.129) (both P<0.05). Conclusions There is a difference in mRNA expression of plasma exosomes between patients with LTBI and ATB. M6PR and RGPD5 may become markers for distinguishing plasma exosomes between LTBI and ATB.