Objective To explore the enhancement effect of caffeine in cisplatin-induced apoptosis of osteosarcoma cell line OS-U2. Methods The osteosarcoma cells were incubated with different concentrations of cisplatin (0.2, 2, 5, 10 and 20?g/L), caffeine (0.2, 2.0mmol/L) and caffeine + cisplatin for 24, 48 and 72 hours. The proliferation of OS-U2 cells was determined by MTT assay, the apoptotic levels were determined by flow cytometry (FCM), and the morphologic changes of apoptotic cells and the positive rates of apoptosis were determined by fluorescence microscopy. Results The proliferation of OS-U2 cells was inhibited, and the apoptotic level was increased when incubated with caffeine and/or cisplatin. There were dose- and time-effect relationships between the lethal effect of cisplatin on osteosarcoma cells and caffeine. Conclusion There is a remarkable enhancement effect of caffeine on cisplatin-induced apoptosis of osteosarcoma cell line OS-U2. It seems that the apoptosis-inducing activity of caffeine may enhance the lethal effect of cisplatin on osteosarcoma cells.

Scaffolds provide a three-dimensional environment that is desirable for inducing and promoting the production of cartilaginous tissue.Ideally the scaffold should:① have directed and controlled degradation;② promote cell viability,differentiation,and extracellular matrix production;③ adhere and integrate with the surrounding native cartilage;④ span and assume the size of the defect;⑤ provide mechanical integrity depending on the defect location;⑥ have no toxicity,no immunogenicity or inflammatory induction;⑦ allow for the diffusion of nutrients and waste products.To date,a wide range of natural and synthetic materials have been investigated as a scaffold for cartilage repair.Based on the morphology and structure,these materials can be divided into hydrogel,sponge,fiber mesh and so on.Natural polymers that have been explored as bioactive scaffolds for cartilage engineering include:alginate,agarose,fibrin,hyaluronic acid,collagen,gelatin,chitosan,chondroitin sulfate,and cellulose.Synthetic polymers currently explored for cartilage repair include poly(?-hydroxy esters),polyethylene glycol,poly(NiPAAm),poly(propylene fumarates),and polyurethanes.

Stimulating factors have been employed to induce,accelerate,and/or enhance cartilage formation.For instance,stimulating factors and other additives may be added to culture media in vitro or incorporated into scaffolds for in vivo delivery to control cellular differentiation and tissue formation.At present,many growth factors and other dissoluble factors such as hyaluronic acid,chondroitin sulfate,and insulin have been used alone or synergistically in cartilage tissue engineering.The application efficacy depends on cell type and culture condition.In addition,gene therapy has emerged as another method of local delivery,where cells can be engineered to over-express bioactive molecules.An additional approach is the introduction of mechanical signals through loading regimes such as hydrostatic or dynamic compression or through the use of bioreactors.Current bioreactors used in cartilage tissue engineering include flat-sheet bioreactor,rotating wall vessel bioreactor and concentric cylinder bioreactor.Bioreactor can improve nutrition transmittability,provide hydrodynamic environment,induce shear stress,and promote production of cartilage specific matrix protein.

Objective To evaluate availability and outcomes of using anterior half of the peroneus longus tendon (AHPLT) in knee ligament reconstruction as an autograft source.Methods From July 2007 to January 2008,100 patients with knee ligament injuries were recruited in this study.There were 33 males and 67 females aging from 16 to 62 years (mean,32.3 years).49 cases had undergone medial patellofemoral ligament reconstruction,19 cases multiligament reconstruction,18 cases double-bundle posterior cruciate ligament (PCL) reconstruction and 14 cases double-bundle anterior cruciate ligament (ACL) reconstruction.AHPLT was used as sole (49 cases) or part (51 cases) of reconstruction materials.One-incision or two-incision striping techniques were adopted to harvest AHPLT.Ligaments were fixed with screws.Post-operative assessments included Kujala knee score,Lysholm knee score,Marx knee score,International Knee Documentation Committee (IKDC) subjective evaluation form and objective evaluation grade,the Foot and Ankle Disability Index (FADI) and the American Orthopedic Foot and Ankle Society (AOFAS) scale.Results 92 cases were followed up for more than 24 months.Postoperative Kujala score,IKDC subjective score,Lysholm score and Marx score were improved significantly in all four groups of patients.According to IKDC objective evaluation grade,the number of patients reaching Grade A (normal) or Grade B (near-normal) in multiligament,PCL and ACL reconstruction were 17,15 and 12,with an excellent rate of 89.5％ (17/19),93.7％ (15/16) and 100％ (12/12),respectively.Preoperative and postoperative AOFAS scores were 97.4±2.0 and 97.2±1.6,respectively,while the FADI scores preoperatively and postoperatively were 96.8±2.2 and 96.9±2.5,respectively.These results had no statistical significance.No signs of peroneal nerve injury or peroneus longus tendon rupture was found.Conclusion It is acceptable to use AHPLT as an autograft due to its feasibility to harvest,good clinical outcome,and low rate of donor site morbidity at a minimum of two-year follow-up.

A new method to measure arterial baroreflex-blood pressure control (ABR-BP) in conscious rats was established in our laboratory. ABR-BP was quantitatively determined by comparing the pressor responses to All before and after the blockade of the efferent pathway of baroreflex. It was found that there was a close relationship between Hood pressure variability and ABR-BP rather than arterial baroreflex-heart period control. ABR-BP was significantly lower in SHR and RVHR than in WKY rats. The lower ABR-BP in SHR was due mainly to the impairment of parasympathetic component

Purpose:To evaluate intraoperative block of abdomial aorta(BAA) in surgery of sacral and pelvic tumors as a useful adjuvant technique.Methods:Of 164 cases of pelvic tumor who underwent resections and hemisections, the procedures in 94 cases were non block of abdominal aorta(NBAA);in 109 cases of sacral tumors various extents of acrectomy was done,53 cases had NBAA,56 cases had BAA.Results:In the pelvic tumor group, the size of tumors for BAA were on the average bigger than NBAA by 0.8 cm, the operation was shortened by 2 h, the dose of hemorrhage decreased by 2 200 ml. The complications of operation were reduced by 6.3%, death rate reduced by 4.6%. In the sacral tumor group, the size of tumors for BAA were on the average bigger than NBAA by 0.7 cm, the operation was shortened by 1 h and 40 min, the dose of hemorrhage decrease by 1 600 ml. The complications of operations were reduced by 23%, death rate reduced by 7.5%.Conclusions:The adoption of BAA adjuvant technique can reduce the intraoperative hemorrhage,the death rate, and the complications of operation, while also shortening the surgical operating time, and is a valuable clinical technique. [

Objective To summarize the experiences of "three holes and one hook in the end(TOE)" laparoscopic cholecystectomy(LC) in 1 260 cases and to investigate the operation procedures,technical points and the prevention of complications.Methods The data of 1 260 patients suffering from chronic calculous cholecystitis,acute calculous cholecystitis,atrophic cholecystitis,gallbladder polyps etc.,who were admitted to this hospital and treated by TOE from March 1999 to March 2008 were included and analysed retrospectively in this study.Results One thousand two hundred and sixty of cases were cured,including 1 252 cases of succeeding LC(99.37%),conversions to open in 8 cases,no death,no bile duct injury,with intraoperative hemorrhage in 3 cases,umbilicus infection in 2 cases,gallbladder fossa hydrops in 3 cases,with operation time for 8-60 min(mean 38.5 min) and hospitalization for 3-7 d(mean 5 d) after surgery.During the follow up of 1 002 cases for 1 to 7 years(mean 3.5 years),there were no complications such as bile fistula,bile duct stricture,residual stones of biliary duct,etc..Conclusion TOE is worthy of application and promotion for the excellent effectiveness,few complications,rapid recovery and safety.

Objective To investigate the safety and feasibility of endovascular stent angioplasty in treating symptomatic stenosis of middle cerebral artery.Methods Endovascular angioplasty with coronary stents was performed in 27 patients with symptomatic stenosis of middle cerebral artery.The clinical results were reviewed and analyzed.Results Of the total 27 patients,successful placement of the coronary stents Was achieved in 24.Angiography immediately after the procedure showed that the stenotic degree of the diseased artery was markedly decreased from preoperative (80±19)%to postoperative (8±4)%,the improvement was very obvious.Percutaneous transcatheter angioplasty had to be employed in two cases because of the failure of stent placement.A mean follow-up period of 18 months was carried out.During the following up period no transient cerebral ischemia attack occurred in 25 patients and no newly-developed cerebral infarction in region fed by the responsible vessels occurred either.Re=irrigation cerebral hemorrhage was seen in one patient,which occurred three hours after the placement of the stent.In one case the placed stent fell off and immigrated into the siphon of internal carotid artery,and the displaced stent Was took out later with a catching apparatus.In another case re-stenosis occurred six months after the stenting.Conclusion Percutaneous endovaacular stent angioplasty is a safe and effective treatment for symptomatic stenosis of middle cerebral artery,although its long-term results need to be further evaluated.

Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 to detect the Treg cells in patients with gastric cancer.Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3.FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7.The mRNA and protein expression of Foxp3 in isolated CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages of Treg cells were more abundant in ascites(t=2.357,P<0.05),TIL(t=6.174,P<0.01) and tumor-draining lymph nodes(t=5.481,P<0.01)of individuals with gastric cancer than that in their blood.There were significant differences in the prevalence of Treg ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.

Objective To investigate the populations of CD+4CD25high Foxp3+ regulatory T cells(Treg cells)and mRNA expression of Foxp3 in peripheral and the marginal region of tumor from patients with gastric cancer,and to evaluate the prevalence of Treg cells from patients with gastric cancer in relation to the TNM stages.Methods PBMC,Ascites,tumor-infiltration lymphocyte and tumor-draining lymph nodes in 47 patients with gastric cancer(TNM I11,Ⅱ18,Ⅲ10,Ⅳ8)and PBMC in 20 normal healthy donors were evaluated for the proportion of Treg cells,as a percentage of the total CD4+ cells,by flow cytometric analysis.Levels of mRNA for Foxp3 were measured witll a real-time quantitative PCR Results The percentage of CD4+CD25high Foxp3+ Treg cells in PBMC for cases of gastric cancer were significantly higher than those for healthv donors(P<0.05).The percentage of Treg cells were more abundant in ascites(P< O.05),1rIL(P<0.01)and tumor-draining lymph nodes(P<0.01)of individuals with gastric cancer than in their blood.And the Foxp3 mean fluorescent intensity (MFI) of Treg cells in TIL and tumor draining lymph nodes with gastric caners were significantly higher than PBMC and ascites(P<0.05).The proportion 0f Treg cell in patients of gastric cancers with stage I,II,Ⅲ,Ⅳ were(5.72±1.95)%,(6.45± 2.23)%,(9.58±3.13)%,(11.70±2.30)%,respectively.There were significant correlation between Drevalenee of Treg cells and disease stages in gastric cancer(r=0.784,P<0.01).Foxp3 mRNA levels were higher in PBMC from the group with gastric cancers than in the group with healthy donors.Foxp3 mRNA levels were correlated with TNM stages(r=0.623,P<0.05).Conclusions Our results suggest that the populations of CD4+CDhigh25 Foxp3+Treg cells and Foxp3 mRNA levels in patients with gastric cancer are signifieantlv higher in comparison to those in control donors.In addition,the expression of Fox：p3 mRNA increased with tumor stage.The increased prevalence of Treg cells may be one of the explanations for impaired cell-mediated immunity in cancer bearing hosts.