To study the changes in serum immunosuppressive acidic protein (IAP) and T lymphocytes rDNA transcription activity in peripheral blood of patients with pancreatic cancer before and after operation. The level of IAP and T lymphocytes rDNA transcription activity in peripheral blood of 58 patients with pancreatic cancer were measured before and after opreration by one directional immunodiffusion test and a cell image analysis of Ag NORs, respectively, and the results were compared with that of healthy controls. The level of IAP was higher while T lymphocytes rDNA transcription activity was lower before operation compared with healthy controls( P 0 05). The levels of IAP and T lymphocytes rDNA transcription activity were closely related to tumor invasion, metastasis and clinical stages (TNM). Serial determination of the levels of IAP and T lymphocytes rDNA transcription activity might serve as an important index for evalution of tumor invasion and metatasis, and also prognosis for the patients with pancreatic cancer

AIM: To observe the realgar induced the apoptosis of eosinophils from bronchoalveolar lavage fluid in asthmatic guinea pigs and investigate the mechanism that realgar treated asthma.METHODS: The morphology of apoptosis of eosinophils was observed by Giemsa staining and electron microscope.The rate of apoptosis of eosinophils was assayed by the flow cytometry.RESULTS: The characteristic changes of the apoptosis in both light microscope and electron microscope were shown after 6 hours treatment of realgar.Flow cytometry showed that the rate of apoptosis of the eosinophils was increased with both increasing realgar concentration and prolonging realgar action time to the cells.CONCLUSION: Realgar promotes the apoptosis of eosinophils from bronchoalveolar lavage fluid in asthmatic guinea pig.Realgar induced the apoptosis of eosinophils is one of the causeses for asthmatic treatment.

Objective To study the role of cytidine-phosphate-guanosine oligodeoxynucleotide(CpG ODN) on the development of Plasmodium liver stage.Methods Plasmodium yoelii BY265 18S rRNA was cloned,and the TaqMan real-time PCR was established on P.yoelii BY265 18S rRNA and mouse GAPDH as quantitative analysis model.The model was tested by the level of liver Plasmodium load with the liver cDNA in BALB/c mice infected by salivary gland sporozoites for 42 hours.Twelve BALB/c mice were randomly divided into CpG group,CpG control group and PBS control group which were injected respectively by ODN1826 30 ?g,ODN1826 control 30 ?g and 0.01 mol/L PBS 200 ?l via vena caudalis.Twenty-four hours later,each mouse was inoculated with 100 sporozoites.Mice were sacrificed in 42 hours after infection,and the liver load of Plasmodium was analyzed by TaqMan real-time PCR.Results The cloned Py BY265 18S rRNA gene showed 98% similarity to Py 17XNL.The quantitative analysis model consisted by 18S rRNA and GAPDH showed positive correlation between the level of liver Plasmodium load and the sporozoite inoculation dose to mice.The Plasmodium load in CpG ODN pre-treated mice was reduced to one fifth of the control group(0.28/1.33)(P

Objective To compare the efficacy and safety of two kinds of homemade sirolimus-eluting stents (Firebird and Excel) for treatment of acute ST segment elevation myocardial infarction (STEMI) in patients who underwent percutaneous coronary intervention (PCI). Methods The 249 consecutive patients with STEMI who underwent PCI were randomly divided into two groups: Excel group (n=136) and Firebird group (n=113). They were followed up for 6-24 months, and coronary angiography was reviewed average 12 months later. The primary endpoints were major adverse cardiac events, including death, reinfarction and target vessel revascularization. The second endpoints included late luminal loss and restenosis 12 months after treatment. Results There were no significant differences in baseline data, coronary arterial lesion before operation, and immediateness condition after PCI between the two groups (all P＞0.05). Within follow-up, there were 2 (1.47%) death cases and 1 (0.88%) death case, 1 (0.74%) and 1 (0.88%) nonfatal myocardial infarction case, 2 (1.47%) and 2 (1.77%) target vessel revascularization cases in the two groups respectively (all P＞0.05). There were no significant differences in late luminal loss of in-stent and in-segment, the rates of in-stent restenosis, in-segment restenosis and stent thrombosis, the in-stent minimal lumen diameter and in-segment minimal lumen diameter between the two groups (all P＞0.05). Conclusions The two kinds of homemade sirolimus-eluting stents may have similar efficacy and safety in patients with STEMI treated with primary PCI.

Aim To investigate the effect of Tetramethylpyrazine (TMP) on the proliferation of airway smooth muscle cells (ASMCs). Methods Primary ASMCs of rats were cultured. The absorbance (A490) value of ASMCs treatment with platelet-derived growth factor (PDGF) in the presence of TMP was detected by MTTto observe the anti-proliferation of TMP. The levels of ERK1/2 and p-ERK1/2 proteins were determined by Western blot.Results In presence of the TMP with different concentrations (12.5,25,50,100 and 200 ?mol?L-1) at 6,12,24,36 and 48 hours,compared with control groups,the average inhibitory rates of cell proliferation in all groups were increased significantly (P

BACKGROUND:How to obtain sufficient autogeneic chondrocytes is a problem which must be answered as soon as possible in both the transplantation of chondrocytes and the development of cartilage engineering.Bone marrow-derived mesenchymal stem cells have the potential of multidirectional differentiation. Under different induced conditions, they can differentiate into multiple tissue cells, such as chondrocyte, osteoblasts , sarcoblast, nerve cells and so on. Insulin-like growth factor-I plays an important role in regulating the formation and development of limb and cartilage.OBJECTIVE:To observe the effect of the insulin-like growth factor-I and culture solution of chondrocyte on inducing bone marrow-derived mesenchymal stem cells to differentiate into chondrocytes in vitro.DESIGN:Open experiment.MATERIALS:This experiment was carried out at the Medical Study Center, Second Hospital Affiliated to Sun Yat-sen University from March to November 2003.Human bone marrow-derived mesenchymal stem cells were harvested from 4 to 6-month old embryos, all of which were from pregnant women who needed to terminate of pregnancy by induction delivery with water bag for health.METHODS:Human embryonic mesenchymal stem cells were isolated with Percoll separating medium. Subsequently, the cells were amplified in vitro,and the expression of surface makers of bone marrow-derived mesenchymal stem cells, such as CD44, CD71, CD34 and CD45 were measured with flow cytometer to identify the cells in our experiment. 100 μg/L insulin-like growth factors and culture solution of chondrocytes were added in the culture medium of bone marrow-derived mesenchymal stem cells of the fourth generation. The morphological changes of induced cells were observed with an inverted microscope. The expression of type Ⅱ cartilage matrix was observed by collogen immunohistochemistry. The proteoglycan level in the cells was detected, too.MAIN OUTCOME MEASURES:Phenotype of bone marrow-derived mesenchymal stem cells was identified through detecting the expressions of CD34, CD44 and CD4.Type Ⅱ collagen immunohistochemistry and the change of cellular ability to secrete proteoglycan after induction were observed to determine whether bone marrow-derived mesenchymal stem cells can differentiate into chondrocytes.RESULTS: ① Being observed under an inverted microscope,the bone marrow-derived mesenchymal stem cells presented a morphology like fibroblasts when they were cultured in vitro. ②Identification of surface antigen of bone marrow-derived mesenchymal stem cells: These cells were detected to have good homogenicity with flow cytometer. The fourth generation of bone marrow-derived stem cells positively expressed CD44, negatively expressed CD34 and CD45, suggesting these cells had the characteristics of bone marrow-derived mesenchymal stem cells. ③Observation of the morphological change of chondrocytes induced by bone marrow-derived mesenchymal stem cells under optical microscope: chondrocyte condition culture solution and insulin-like growth factors-Ⅰ were added to co-culture.During the process of culture, bone marrow-derived mesenchymal stem cells were seen to have a shape of round gradually. Fifteen days later, some cells presented a shape of short-shuttle or polygon with short mutations,which were the shape characteristics of chondrocytes. ④Immunohistochemical staining of Type Ⅱ collagen: In the insulin-like growth factor group,72.5% cells had many brown granules in cytoplasm, which were weakly positive or strongly positive expression of Type Ⅱ collagen. In the control group, bone marrow-derived mesenchymal stem cells was negative expression of type Ⅱ collagen on the 15th day. ⑤ Measurement of proteoglycan level: After co-culture with insulin-like growth factor-Ⅰ and chondrocyte culture solution for 15 days, proteoglycan was higher in the cells of co-culture group [ (8.92±0.91) μg/L ] than in culture group [(2.56±0.26) μg/L,P ＜ 0.05], but lower than in the chondrocyte group[(13.69±1.51) μg/L, P＜ 0.05].CONCLUSION:Insulin-like growth factor-Ⅰ and chondrocyte culture solution can induce bone marrow-derived mesenchymal stem cells to differentiate into chondrocytes.

Objective To determine whether field triage would reduce median contact-to-device ( C2D ) time in patients with ST-segment elevation acute myocardial infarction ( STEMI ) . Methods Consecutive patients with STEMI underwent primary percutaneous coronary intervention( PCI) from March 2010 to February 2014 in Shanghai Pudong Gongli Hospital were analyzed. Patients were divided into two groups. A total of 121 patients were admitted by field triage and 101 patients by non-field triage. The primary study point was C2D time and the study points secondary included ( door-to-balloor, D2B) time, peak Troponin I ( TnI) levels, hospital mortality and 30 days follow-up mortality. Results Baseline and procedural characteristics between the two groups were comparable. Comparing to non-field triage group, the C2D time was reduced [(92. 0 ± 56. 0)min vs. (131. 0 ± 61. 0)min,P﹤0. 01]. The D2B time was lower in the field triage group vs. the non-field triage group [(55. 0 ±26. 0)min vs. (96. 0 ±31. 0)min,P﹤0. 01]. The percentage of patients with C2D time less than 90 minutes increased significantly from 85. 1% to 98. 3%( P﹤0. 01 ) in the field triage group. Peak TnI level was significantly reduced in the field triage group [(23. 5 ±22. 0) μg/L vs. (43. 5 ± 39. 0) μg/L,P﹤0. 01]. In-hospital mortality and 30 days follow-up mortality did not significantly differ between the 2 groups (3. 3% and 3. 0%, P=0. 885;3. 3% and 5. 0%, P=0. 544, respectively). Conclusions In STEMI patients, field triage was associated with significantly reduced C2D and D2B times.

AIM:To investigate the role of canonical transient receptor potential channel 1 ( TRPC1 ) in the migration of human bronchial epithelial cells (16HBE) induced by transforming growth factor-β1 ( TGF-β1).METH-ODS:Silencing of TRPC1 gene expression was performed by siRNA.The cell activity and apoptosis were measured by CCK-8 assay and flow cytometry, respectively.The migration and invasion abilities of the 16HBE cells were detected by wound-healing assay and Transwell assay.The protein expression of E-cadherin and vimentin was determined by Western blot.RESULTS:TGF-β1 treatment significantly enhanced the cell migration distance compared with control groups ( P<0.01 ) .The results of CCK-8 assay and flow cytometry indicated that there were no significant difference in proliferation and apoptosis among TRPC1 siRNA group, TGF-β1 group and control group (P>0.05).The results of wound-healing and Tr-answell assays showed that migration and invasion abilities in TRPC1 siRNA +TGF-β1 group were markedly suppressed compared with TGF-β1 group (P<0.01).The protein expression of E-cadherin and vimentin induced by TGF-β1 was in-hibited by TRPC1 silencing compared with TGF-β1 group (P<0.05).CONCLUSION:TRPC1 is involved in the migra-tion of human bronchial epithelial cells induced by TGF-β1 through regulating the protein expression of E-cadherin and vim-entin.

[ ABSTRACT] AIM:To investigate the role of canonical transient receptor potential channel 1 ( TRPC1 ) in the epithelial-mesenchymal transition ( EMT) of human bronchial epithelial ( HBE) cells induced by transforming growth fac-tor-β1 (TGF-β1).METHODS:EMT of 16HBE cells induced by TGF-β1 were identified by microscopy, immunofluores-cence and Western blotting.Immunofluorescence, real-time PCR and Western blotting were applied to detect the mRNA and the protein expression of TRPC1 in the 16HBE cells.The influence of SKF96365 (a TRPC1 blocker) and siRNA-me-diated silencing of TRPC1 on the EMT of the 16HBE cells were detected by microscopy and Western blotting.RESULTS:Treatment with TGF-β1 induced significant morphological changes of the 16HBE cells.Exposure to TGF-β1 decreased the expression of E-cadherin protein (P<0.01) and increased the expression of α-SMA protein (P<0.05) in the 16HBE cells.Immunofluorescence observation indicated that TRPC1 expression in the 16HBE cells was positive.The expression of TRPC1 at mRNA and protein levels was significantly increased in the 16HBE cells after stimulation with TGF-β1 ( P<0.05).The morphological changes of the 16HBE cells induced by TGF-β1 were inhibited by SKF96365 and TRPC1 silen-cing compared with TGF-β1 group.The protein expression of E-cadherin andα-SMA induced by TGF-β1 were inhibited by SKF96365 and TRPC1 silencing compared with TGF-β1 group (P<0.05).CONCLUSION:TGF-β1 induces EMT with the mechanism of up-regulating TRPC1 in human bronchial epithelial cells.

Objective To establish an aging model of mesenchymal stem cells (MSCs) and to investigate aging related biological mechanism for the purpose of studying the senesence of MSCs .Methods MSCs were separated and purified from human placenta, and the cells of the third passage(P3-MSCs) were cultured in the medium for 2 hours, then 100,200 and 300 μmol/L hydrogen peroxide ( H2 O2 ) was added to the cells for 2 hours to establish the MSCs aging model in vitro. Biological characteristics of aging MSCs were evaluated by cell cycle assay and senescence associated β-galactosidase staining.The expression of p16,p21 and p53 genes was further measured using quantitative real-time PCR (RT-PCR).Re-sults Compared with the control , the number of MSCs treated with 200μmol/L H2 O2 for 2 hours was significantly decreased and the cells displayed less adipogenic ,osteogenic and chondrogenic differentiation .Moreover ,after exposure to 200 μmol/L H2 O2 , the majority of the cells were in the G 0/G1 phase as showed by cell cycle analysis .The percentage of senescence-associated β-galactosidase-positive cells was increased , and the expression of p 16 , p21 and p53 mRNA and protein was significantly increased.Conclusion The results of this study has demonstrated that the H 2 O2 (200 μmol/L) can be used to establish the aging model of MSCs in vitro, and the cellular phenotypic alteration may attribute to the cell cycle associated gene expression (p16, p21, and p53).