Abstract:Objective:To establish a stable glycine receptor alpha 1(GlyRα1)-expressed HEK293 cell line. Methods:The pcDNA3.1-GlyRα1 plasmid was constructed and transfected into HEK293 cells with lipofectin. The stable transfectants were screened by G418. The mRNA expression of GlyRα1 was identified using RT-PCR. The protein expression was analyzed by Western blot. Results:Selected by G418, 4 out of 7 transfected cell lines showed high expression level of GlyRα1, as demonstrated by RT-PCR and Western blot analysis. No or low expression of GlyRα1 was shown in other 3 cell lines. Conclusion:A HEK293 cell line stably expressing GlyRα1 was constructed successfully.

Abstract:Objective:To construct a EGFP(enhanced green fluorescent protein)-labled eukaryotic expression recombinant plasmid of heNOS(human endothelial nitric oxide synthase)gene. Methods:According to the gene of heNOS and the multiple clone site of the expression vector pEGFP-N1 plasmid,a specific pair of primers were designed and synthesized. PCR amplification of heNOS gene from the pBluescript Ⅱ SK-heNOS plasmid was performed,and an approximate 3.6 kbp objective fragment was achieved. The objective fragment was inserted into the downstream of the carrier promoter and combined with the report gene EGFP. The recombinant plasmid was verified by PCR of the bacterium liquid,restriction enzyme digestion and gene sequencing. Results:The heNOS fragment from pBluescript Ⅱ SK-heNOS plasmid were amplified and inserted into the pEGFP-N1 carrier. The enzyme digesting spectra of the recombinant plasmid was correct,and the heNOS gene sequence was completely in accordance with the sequence provided by the Genbank. Conclusion:The pEGFP-N1-heNOS eukaryotic expression recombinant plasmid were successfully constructed.

Abstract:objective:To establish an auto-fluorescent K562 cell model using recombinant retroviral vector system. Methods:The full length cDNA of EGFP gene was inserted into the pCMV-hyg to construct recombinant retroviral plasmid pCMV-EGFP-hyg. The retrovirus containing EGFP was produced by co-transfection of T293 cell line with pCMV-EGFP-hyg, pCMV-G and pCMV-GP. The K562 cells expressing EGFP(EGFP-K562) were obtained by co-culture with recombinant retrovirus-producing T293 cells and Hyg selection. EGFP expression was examined with flow cytometry(FCM) and fluorescence microscopy. To show the application of EGFP-K562 in laboratory research, NK activity assay using EGFP-K562 as target cells was carried out in a series of effector(E) to target(T) cell ratio. Results:Recombined retrovirus containing EGFP was successfully constructed with high titers through co-transfection of T293 cells with pCMV-EGFP-hyg, pCMV-G and pCMV-GP. The highest titer of recombinant virus was 1.5×106CFU/mL when harvested at time point of 48 hours. Strong fluorescent in EGFP-K562 cells were developed and retained without any change during up to 20 passages of cell culture. More than 98% EGFP-K562 was achieved after selection with Hyg. NK activity assay employing EGFP-K562 as target cells showed peripheral blood mononuclear cells(PMNC) exhibited a different cytotoxicity with different E ∶ T ratio and incubation time. The optimal condition of NK activity assay was at a ratio of 40 ∶ 1 between E and T with 4 h incubation. Conclusion:Auto-fluorescent K562 cells was stably established which might provide a novel cell models in the experimental studies.

Abstract:Objective:To investigate the effects and mechanisms of free fatty acid(FFA) on 3T3-L1 preadipocyte differentiation. Methods:11-茁-HSD1-SiRNA expression plasmid(pGCsilencerTM H1/TetO1-11-茁-HSD1) was constructed and transfected into 3T3-L1 cells. 100 μg/ml FFA was added to the 3T3-L1 cells. Levels of adipogenesis-associated markers such as Peroxisome proliferator-activated receptorγ(PPARγ), CCAAT enhancer binding protein α(C/EBPα), lipoprotein lipase(LPL), and fatty acid synthetase(FAS) were measured. Results:When the cells treated with FFA, mRNA expressions of PPARγ,C/EBPα,LPL and FAS were up-regulated and the expression of 11-茁-HSD1 also increased. When 11-茁-HSD1 was silenced, the levels of the markers were down-regulated. Conclusion:FFA and 11-茁-HSD1 could promote adipocyte differentiation, and the effect of FFA on adipogenesis carried out through increasing the expression of 11-茁-HSD1.

Abstract:Objective:To research the relationship of Sulindac and human’s γδT cells on preventing digestive tract’s tumor. Methods:Use the isopentenyl pyrophosphate method to amplify human peripheral blood γδT cells in vitro. After different concentration’s Sulindac to induce γδT and SGC-7901 cells, flow cytometry was used to detect the cell surface marker of γδT cells before and after cultivation and the apoptosis percentage of γδT cells and SGC-7901 by Sulindac. Meanwhile, the killing and wounding activity of γδT cells was measured by lactate dehydrogenase method. Results:γδT cells were amplified from 4.21%to 70.35% in 10 days, CD44 up to 94.0%. when Sulindac’s density is 100 -滋mol/L, the growth inhibition ratio of γδT cells that up to 42.1% was notably higher than gastric carcinoma cell SGC-7901’s growth inhibition ratio(7.4%). After γδT cells and SGC-7901 induced by different concentration of Sulindac for 24 h, the cytotoxicity to SGC-7901 is the highest under 12.5 -滋mol/L Sulindac. The apoptosis percentage of γδT cells(52.71%) was significantly higher than SGC-7901’s apoptosis percentage(7.88%) after induced by Sulindac(100 -滋mol/L) for 24 h. Conclusion:Sulindac can increase the effect of γδT cells killing tumor cells in clinical routine drug concentration, while it can strikingly inhibit the reproductive and killing activity of γδT cells if its concentration higher than routine dose, but this inhibition effect is not obvious for gastric carcinoma cell. This result may offer an experimental base for applying clinical NSAIDs to prevent digestive tract’s tumor.

Abstract:Objective:To study the effect of IL-6 in SLE serum on the differentiation and maturation of dendritic cells(DCs)derived from cord blood CD34+ hematopoietic precursor cells(HPCs). Methods:CD34+HPCs were isolated and purified from cord blood by magnetic cell sorting system(MACS), and cultured with GM-CSF+IL-4+TNF-α. Normal serum or SLE serum with high level of IL-6 was added into culture medium respectively. The phenotypes of DCs(HLA-DR,CD80,CD86,CD83 and CD1a) were analyzed by FCM. Their abilities of stimulating allogenic T lymphocytes proliferation were examined by CCK-8, and ELISA was used to assess the IL-12, IL-10 and IFN-γ concentration in the culture supernatant. Results:The serum level of IL-6 was significantly higher in SLE patients than in controls. When cultured in SLE serum with high level of IL-6, CD34+HPCs could be induced to differentiate into DCs which produced little IL-10, little IFN-γ and lower IL-12, but expressed higher levels of HLA-DR, CD80 and CD86, and showed increased abilities of stimulating allogenic T lymphocytes proliferation compared with those cultured in normal serum. Conclusion: The high level of IL-6 in SLE serum has abnormal impacts on the differentiation and maturation of DCs and alters their characteristics, which may be involved in the initiation and maintenance of SLE.

Abstract:Objective:To explore the generation of mature dendritic cells(DCs) induced from human peripheral blood monocytes in vitro. Methods:Monocytes were isolated from normal human peripheral blood mononuclear cells(PBMCs)and were separated into five groups after cultured with GM-CSF and IL-4 5 days later. The control group were added only GM-CSF and IL-4. One group were added polyriboinosinic polyribocytidylic acid[Poly(I:C)], one group were added calcium ionophore(CI),one group were added Poly(I:C) and CI, and the other group were added TNF-α. The morphology of cells was observed by microscope. The surface antigens of the induced cells were analyzed by fluorescence-activated cell sorting(FACS). T cell proliferating activity was determined by allo-MLR(mixed lymphocyte reaction) in vitro. Results:Large amount of mature DCs could be obtained from monocytes by culture in presence of GM-CSF,IL-4,Poly(I:C) and CI. DC expressed high level of CD83, CD80, CD86 and CD40, and were potent to stimulate the proliferation of allo-lymphocytes. Conclusion:The phenotypes and functions of DC generated from PBMC by the treatment of cytokine combination can be improved by calcium ionophore and Poly(I:C).

Abstract:Objective:To investigate the expression of axon guidance cue slit-2 mRNA, and the ethology and morphology changes in different phases of spinal cord injury(SCI) development. Methods:Forty Sprague-Dawley rats were randomly divided into normal group, sham operation group and operation group which include 6 subgroups(1,3,7,14,21,28 days). The spinal cord hemisection injury model was made by cutting the posterior right side spinal cord. Function changes in the right posterior limb were evaluated by BBB method, the general morphology changes were observed by HE stain, and the expression of slit-2 mRNA was detected through in situ hybridization methods. The positive cells were analyzed by a computer image analysis system. Results:The limb function of experimental rats significantly recovered from 7 d to 28 d. The morphology gradually changed from local hematoma to inflammatory cell infiltrate, scar formation and vacuolar degeneration of the distal as well as the proximal spinal cord tissue. The in situ hybridization results indicated that expression of slit-2 mRNA in the angulus anterior was low in either normal or sham operation group, but significantly increased at 3 d(P < 0.01) in the operation group, then it decreased significantly at 7 d and 14 d(P < 0.05), and continued expression until 28 d. Meanwhile, in 7 d group, the expression of slit-2 mRNA was obviously rising up in the reactive hyperplasia cells around the lesion incision. Conclusion:The injury of adult mammalian spinal cord is a dynamic complex course. The axon guidance cue slit may act as a inhibitory role to repel axon regeneration via mediating migration of inflammatory cells, interacting with several neurogen or cues, participating formation of the scar and other probable mechanisms.

Abstract:Objective:To observe the homing capacity of bone mesenchymal stem cells(BMSCs) to rat with acute hepatic failure(AHF). Methods:BMSCs were isolated from rat bone marrow with the method of density gradient centrifugation and stick-to-wall screening,and the surface markers were identified with flow cytometry after amplification in vitro. BMSCs were labeled with Bromodeoxyuridine(BrdU). AHF animals were induced by D-galactosamine(D-gal). Transplantation of labeled BMSCs(1~1.5×106) were performed on the AHF rats(group A)/normal rats(group C),another AHF group(group B) were injected with sodium chloride without BMSCs as a control to group A. Then the mortality of the rats and liver function recovery status were observed. Two weeks after cells transplantation, BMSCs traced with BrdU were detected in the livers by immunohistochemistry. Results:The mortality and status of liver function were not significantly different between A and B group. Both A and C group could find BrdU+ cells, and the number and the disposition of BrdU+ cells in A group was much more than C group-s,showing significant difference. Conclusion:BMSCs have capacity of homing to the liver of rat in normal or in injury . But the number of BMSCs homing to liver might be related to the extent of liver injury.

Abstract:Objective:To explore the effect of the human mesenchymal stem cells(hMSCs) transplantation on treating experimental autoimmune encephalomyelitis(EAE) rats and the status of transplanted hMSCs in the brain tissue of EAE rats. Methods:hMSCs were isolated from human adult bone marrow. EAE rat models were made, and the human mesenchymal stem cells were transplanted into the rat tail vein 10,15 and 21 days after immunization. The neurological signs were undertaken in all the groups every day after immunization. Rats were sacrificed at different time. The differentiation and migration of the human mesenchymal stem cells in vivo,and the number and status of the demyelinated foci were checked by electronic microscope and immunohistology. Results:Rats that received transplantation had a significant improvement compared with the control group in the observation of electronic microscope. Twenty days later, rats that received transplantation 10 and 15 days after immunization had a significant improvement of clinical outcome compared with the control group(P < 0.05). The number of demyelinated foci decreased significantly(P < 0.05). The human mesenchymal stem cells were induced to differentiate into astrocytes, oligodendrocytes and neurons in vivo. Immunohistological result confirmed that the transplanted stem cells migrated to the demyelinated foci. Conclusion:The human mesenchymal stem cells differentiated into three main types of neural cells by the influcence of the EAE microenvironment signals.Human mesenchymal stem cells transplantation could effectively improve the neurologic function of rats, and decrease demyelinated foci number.

Abstract:Objective:To establish an efficient and stable method for simultaneous isolation and culture of murine mesenchymal stem cells and endothelial progenitor cells from bone marrow. Methods:The mononuclear cells were isolated from murine bone marrow and cultured for 2 days. Then the adherent cells were cultured in LG-DMEM with 10% preselected FBS for MSC and the suspending cells were cultured in EGM-2 MV for EPC, respectively. Osteogenic and adipogenic induction was performed on MSCs. The percentage of MSCs was analyzed by flow cytometry(FCM). EPCs were stained with Dil-ac-LDL and FITC-UEA-1, and the expression of vWF and CD31 was also assessed by immunohistochemistry. Results:The early attached cells exhibited clonal morphology as early as 48 hours, which became 80% confluent within 1 week. MSCs could differentiate into osteocytes and adipocytes after induction. The expression of CD29,CD34,CD45 and CD90 were(93.86 ± 1.12)%,(0.48 ± 0.38)%,(1.89 ± 1.49)%,(94.11 ± 3.32)%, respectively. Cells cultured in EGM-2 MV became confluent after 2 weeks,(75.2 ± 4.5)% of which were double positive for Dil-ac-LDL and FITC-UEA-1 staining. The percentages of vWF+ and CD31+ cells were (55.7 ± 4.7)% and (52.5 ± 3.6)%, respectively. Conclusion:An efficient, stable and replicable method for simultaneous isolation and culture of MSCs and EPCs from a single mouse was established.

Abstract:Objective:To elucidate the adaptive changes on the morphology and proliferation of pancreatic islet during pregnancy in rats. Methods:Rat pancreatic tissues of each group were double-stained by 5-bromodeoxyuridine(BrdU) and anti-insulin antibody by immunohistochemistry technique to measure the changes of islet morphology and quantity during pregnancy at day 10.5,12.5, 14.5,18.5 and 20.5. Results:Compared with the normal control group, the average photodensity of insulin-expression positive zone, insulin relative concentration(IRC), insulin-expression positive cell density(PCD) and proliferation index(PI)of pancreatic islet were all significantly increased on day 12.5, rose continuously to a peak at day 14.5 of pregnancy, and then gradually returned to levels as control. In addition, areas of insulin-expression positive cells were also much greater during pregnancy during day 14.5 to day 20.5 than in non-pregnant rats. Conclusion:Both pancreatic islet -茁 cell proliferation and insulin-expression concentration became more significantly increased than sex-matched virgin control rats from day 12.5, peaked at day 14.5 of pregnancy,and lasted to late pregnancy(day 20.5).

Abstract:Objective:To compare the diagnostic value of serum tartrate-resistant acid phosphatase 5b(Tracp5b), cancer antigen 153(CA-153) and carcino embryonic antigen(CEA) in patients with bone metastases of breast cancer. Methods:The serum levels of Tracp5b, CA-153 and CEA were determined by enzyme-linked immunosorbent assay and electrochemiluminescence in 26 breast cancer patients without bone metastases, 32 breast cancer patients with bone metastases and 18 healthy female adults. Results:The serum levels of Tracp5b in breast cancer patients with bone metastases were significantly higher than those in other two groups(P < 0.05). The sensitivity and specificity of Tracp5b to bone metastases was 78.13% and 86.36%. The sensitivity and specificity of CA-153 to bone metastases was 37.50% and 77.27%, and those of CEA were 21.88% and 84.09%, respectively. The sensitivity of Tracp5b were significantly higher than those of CA-153 and CEA(P < 0.05). Conclusion:Tracp5b is a useful serum marker to diagnose bone metastases in breast cancer patients.

Abstract:Objective:To compare the antitumor effect of paclitaxel-loaded liposomes with paclitaxel on S180 ascites-tumour bearing mice. Methods:The paclitaxel-loaded liposomes were prepared by reverse evaporation and sonication techniques. The particle diameters were measured by dynamic light scattering(DLS) measurement and the entrapment efficiency was measured by the reverse-phase high-pressure liquid chromatograph(RP-HPLC) method. Twenty Kunming mice bearing ascites tumor xenografts were distributed into four groups at random:the control,empty-liposomes,free paclitaxel group and the paclitaxel-loaded liposomes. Treatments were started after 24 hours of implantation. Two groups bearing S180 xenograft were treated i.p. with 10 mg/kg of paclitaxel(free drug or liposome encapsulated). The other two groups were given i.p. with saline or empty-liposomes. All the treatments were given 4 times and the interval was 72 hours. Results:The average diameter of the paclitaxel-loaded liposome was 282.4 ± 8.94 nm. The entrapment efficiency was 91%. There were resembly much ascites in the three groups of the control, empty-liposomes and free paclitaxel. While in the group of paclitaxel-loaded liposomes,there was little ascites. In addition,there were dramatically little metastasis in abdominal cavity with the group of the paclitaxel-loaded liposomes other than the remain three groups. Conclusion:Ascites and metastasis in abdominal cavity could be inhibited by paclitaxel-loaded liposomes to S180 ascites-tumour bearing mice remarkly.

Abstract:Objective:To study the changes and clinical significance of serum interleukin-6(IL-6), C-reactive protein(CRP),von willebrang factor(VWF) and plasma P-selectin(Ps) in pigs in different stages after left atrial appendage transcatheter occlusion. Methods:Occlusion group with 12 shuzhong pigs and control group with 5 pigs were used in the experiment. The changes of serum IL-6,CRP,VWF and plasma Ps were measured by enzyme linked immunosorbent assay(ELISA) before the operation, immediately at the operation,and 1,3 month after the operation. Results:In occlusion group, the levels of serum IL-6,CRP,VWF and Ps were higher at immediate time after occlusion than those of before occlusion(P < 0.05), but the levels of them at 1 month and 3 month after the occlusion were as high as those of before occlusion(P > 0.05). In control group, the levels of serum IL-6,CRP,VWF and Ps were higher at immediate time after operation than those of before the left atrial appendage transcatheter occlusion(P < 0.05). There were no significant difference in the levels of serum IL-6,CRP, VWF and Ps in different stages after the operation between the occlusion group and the control group(P > 0.05). Conclusion:Levels of serum IL-6, CRP, VWF and Ps temporarily increased at immediate time after the operation. This observation showed the satisfactory biocompatibility, feasibility and safety of the domestic nitinol left atrial appendage occluder.

Abstract:Objective:To detect the expression of placenta growth factor(PlGF) in maternal serum and urine from preeclampsia (PE), and to identify whether the level of PlGF in urine is significantly correlative with that in serum. Methods:Women enrolled prospectively in following groups: healthy pregnant control(n = 20) and PE patients, which contained mild preeclampsia(n = 20)-severe preeclampsia(n = 15). The serum and urinary PlGF levels were measured by enzyme linked immunoassay(ELISA). Results: The serum PlGF levels in severe preeclampsia group(23.40 ± 9.88)pg/ml were significantly lower than that of normal pregnancies(44.04 ± 19.26)pg/ml and mild preeclampsia group(36.13 ± 17.29)pg/ml. The urinary PlGF level in severe preeclampsia group(587.20 ± 381.86)pg/mmol was also significantly lower than that in normal pregnancies(961.65 ± 584.58)pg/mmol and mild pre-eclampsia group(939.20 ± 474.94)pg/mmol. The urinary PlGF level was significantly correlative with that in serum(r = 0.713). Conclusion:Severe preeclampsia is associated with the low serum and urinary placenta growth factor levels in late pregnancy. Urinary PlGF level is significantly correlative with serum PlGF level. The level of PlGF in urine may be used to study the connection between PE and PlGF instead of serum PlGF level and offer a more convenient method to make a diagnosis, which may guide the clinical treatment of PE.

Abstract:Objective:To investigate the expression of N-acetyl-glucosamine-6-O-sulfotransferase,an important enzyme in synthetic enzyme for L-selectin ligant in the endometrium from the patients with carcinoma of endometrium and normal women. Methods:Reverse transcription polymerase chain reaction(RT-PCR) method was used to detect and quantify N-acetyl-glucosamine-6-O-sulfotransferase mRNA expression levels in the endometrium of 16 patients with carcinoma of endometrium which were recruited as study group and 16 healthy women as the control. Results:The expression of N-acetyl-glucosamine-6-O-sulfotransferase was observed in normal, but the expression level in study group was significantly higher than that in control group(0.87 ± 0.08 vs 0.45 ± 0.11,P < 0.001), higher than that in paraneoplastic endometrium of the patients with carcinoma(0.87 ± 0.08 vs 0.49 ± 0.12,P < 0.001). Conclusion:The higher expression of N-acetyl-glucosamine-6-O-sulfotransferase in the endometrium from the patients with carcinoma of endometrium may indicate a close correlation with the development and metastasis of cancer cells.

Abstract:Objective:To explore the role of β-catenin in differential diagnosis between mesenteric fibromatosis and other mesenchymal tumors of gastrointestinal tract. Methods:The expressions of β-catenin, vimentin, smooth muscle actin(SMA), desmin, CD117, CD34 and S-100 in mesenteric fibromatosis, gastrointestinal stromal tumor(GIST), leiomyoma, inflammatory myofibroblastic tumor and nonspecific fibrous hyperplasia were studied by immunohistochemical method. Results:All 5 examples of mesenteric fibromatosis displayed strong nuclear β-catenin staining. All other lesions tested(GIST,leiomyoma,inflammatory myofibroblastic tumor and nonspecific fibrous hyperplasia) lacked nuclear labeling for β-catenin. All tumors showed diffuse strong positive for vimentin. The expressions of CD117 and CD34 were mainly seen in GIST. The expressions of SMA and desmin were mainly seen in leiomyoma. S-100 protein slight staining could be found in few mesenteric fibromatosis, GIST and leiomyoma. Conclusion:Nuclear β-catenin staining is specific and responsive to mesenteric fibromatosis. β-catenin could probably serve as a useful diagnostic and differential diagnostic tool of mesenteric fibromatosis.

Abstract:Objective:To evaluate the effects of ischemic preconditioning(IPC) on the signal transduction pathways of liver regeneration after partial liver transplantation in rat models. Methods:Male Lewis rats were used as donors and recipients of 50% orthotopic liver transplantations. 120 rats were divided into two groups randomly, control group(C) and ischaemic preconditioning treated group(IPC). IPC group was performed by clamping of the portal vein and hepatic artery of the donor for 10 minutes followed by reperfusion for 15 minutes before harvesting. The survival rate of both group were calculated each day up to 7 days. At 2,6,24, 48 h after reperfusion, serum Alanine Aminotransferase(ALT) of both group were measured, graft samples were obtained to determine the hepatic levels of TNF-α,IL-6 and PCNA. The expression of proteins cyclin D1,Cdk4,STAT3 and P-STAT3 were detected by Western blot. Results:After liver transplantation there was no significant difference between both groups on the survival rate. The level of serum ALT at 2 h and 6 h of IPC group lower than the control group. The hepatic levels of TNF-α of IPC group were higher than the control at 6 h. As to the IL-6, group C attenuated at all time spots. The activity of PCNA in IPC group increased significantly compared with the control as showed by Western blot. Cyclin D1 were expressed intensively in IPC group. The results of P-STAT3 were similarly to the Cyclin D1. Cdk4 were expressed intensively in IPC group. There was no difference between both group on STAT3. Conclusion:After 50%-size-graft liver transplantation, IPC can enhance the liver regeneration pathways induced by IL-6 and TNF-α,thus improve the ability of liver regeneration.

Abstract:Objective:To identify the ligustrazine effect on asthmatic mice serum stem cell factor(SCF) expression and its effects on inhibiting bronchi inflammation. Methods:Twenty-four male BALB/c mice were randomly divided into four groups:the normal control group(A), the asthmatic model group(B),the low-dose ligustrazine group(C,40 mg/kg) and large-dose ligustrazine group(D,80 mg/kg). There were 6 mice in each group. The bronchi pathological changes were detected by HE dyeing and the SCF content was examined by ELISA. Results:The content of SCF in mice in asthmatic model group[(114.9 ± 27.3)pmol/L] is higher than that in normal control group[(48.6 ± 11.2)pmol/L](P < 0.01). The SCF contents in low-dose ligustrazine group[(70.6 ± 7.9)pmol/L] and in large-dose ligustrazine group[(51.4 ± 8.1)pmol/L] are both lower than that in the asthmatic model group(P < 0.01). In the asthmatic group,the numbers of eosinophils and lymphocytes,the thicknesses of WA/Pi and ASM/Pi were(32.6 ± 4.5)/mm2,(1.196 ± 0.111)mm,(0.292 ± 0.027)mm,respectively;and those of the normal group were(6.6 ± 1.2)/mm2,(0.571 ± 0.057)mm,(0.139 ± 0.014)mm,respectively;there were significant difference between the two groups(all P < 0.01), The numbers of eosinophils and lymphocytes,the thicknesses of WA/Pi and ASM/Pi of the low-dose ligustrazine group were(26.0 ± 2.5)/mm2,(0.949 ± 0.105)mm,(0.243 ± 0.027)mm, respectively. They were lower than those in the asthmatic group(all P < 0.05). The numbers of eosinophils and lymphocytes,the thicknesses of WA/Pi and ASM/Pi of the large-dose ligustrazine group were(12.1 ± 1.6)/mm2,(0.875 ± 0.111)mm,(0.178 ± 0.023)mm,respectively. They were significantly lower than those in the asthmatic group(all P < 0.01). Conclusion:Ligustrazine may control the mice bronchi inflammation by inhibiting the SCF expression.

Abstract:Objective:To explore the anabolic effect of intermittently adminstrated PTH on rat osteoblast cells(ROBs). Methods:The ROBs divided into two groups:①Control(Ctr),the cells were cultured without PTH in an incubation cycle(24 h);②Intermittent expoure to PTH(Itm), the cells were cultured with PTH at the first 6 h,but without PTH at the subsequent 18 h. Cellular proteins at 24 h were disolved and seperated in two-dimensional gel electrophoresis(2-DE). Two of the dramatically upregulated proteins were detected by matrix assisted laser desorption/ionization time-of-flight mass spectrometry(MADI-TOF-MS). Results:The two upregulated proteins were β-actin and vimentin. Both of them were cytoskeletal proteins. Conclusion:β-actin and vimentin may play a role in the anabolic effect of intermittent PTH on rat osteoblast cells.

Abstract:Objective:To investigate the relationship between single nucleotide polymorphisms(SNP) of adiponectin gene(SNP+ 276G/T) and coronary artery disease in non-diabetic Chinese han population. Methods:The single nucleotide polymorphisms(SNP) at positions 276 in adiponectin gene were determined by polymerase chain reaction-restriction fragment length polymorphisms(PCR-RFLP) assay in persons of Han nationality in Nanjing area, including 135 patients with coronary artery disease(CAD group) and 131 normal controls(control group). Results:Adjusted OR of G/G genotype is 3.16(P < 0.05); adjusted OR of G/G+G/T is 2.69,(P < 0.05). Conclusion:The distribution of SNP+276 genotypes were significantly different between CAD group and control group. And G/G genotype appeared more frequently in patients with coronary artery disease than normal population, which suggests that adiponectin gene seems to closely associate with coronary artery disease in Chinese han population.

Abstract:Objective:To evaluate the effect of percutaneous balloon valvuloplasty(PBMV) in the elderly cases and study the adaptation and management with PBMV in these cases. Methods:This was a prospective single center clinical trial. A total of 56 cases whose age ≥60 years old were performed with modify Inoue PBMV. The hemodynamics and echocardiogram were used to evaluate the left atrial pressure(LAP), mitral valve area(MVA), mean gradient across the mitral valve(MVG), left atrial diameter(LAD) before procedure and immediately after procedure. The echocardiography assessment items before and one year after procedure included the MVA, MVG and LAD. Results:54 cases had successful operation in 56 paitents. 2 cases were failure. LAP from(22.5 ± 6.2)mmHg(1 mmHg = 0.133 kPa) before the operation decrease to(13.8 ± 4.6)mmHg immediately after the operation,(P < 0.01). MVA from(1.05 ± 0.21)cm2 before operation increase to(1.78 ± 0.22)cm2 immediately after operation(P < 0.01). MVG from(18.3 ± 5.2)mmHg before operation decrease to(10.6 ± 4.2)mmHg immediately after operation(P < 0.01). LAD from(4.71 ± 0.73)cm decrease to(4.39 ± 0.48)cm(P < 0.01) one year after operation. Restenosis detected by echocardiography was seen in 3 cases. One case died in mitral valve replacement performed. The echocardiography one year follow-up result showed that the effect of PBMV was constant. Conclusion:PBMV may be performed safely and successfully in old patients with mitral valve calcification and high risk, who can not afford mitral valve replacement.

Abstract:Objective:To study the protective effect and mechanism of anisodamine on cardiopulmonary bypass(CPB) induced lung injury. Methods:According to the medicine dose and administer time,twenty four patients with congenital heart defect (CHD) were divided into four groups. Control group, group Ⅰ(anisodamine 2 mg/kg, priming solution),group Ⅱ(anisodamine 2 mg/kg,main pulmonary artery), group Ⅲ(anisodamine 1mg/kg,priming solution + anisodamine 2 mg/kg,main pulmonary artery). Neutrophil,CD11b and TNF-a were measured at five time points of the operation. Little lung tissue was cut out for detecting NF-κB after the operation. Blood gas analysis were also arranged. Results:The level of neutrophil,CD11b and TNF-α was reduced in group Ⅰ, group Ⅱ and group Ⅲ compared with the control group, especially in group Ⅱ and group Ⅲ. The results of NF-κB was conformity with the former. Certinary blood gas analysis changed better. Conclusion:CPB induced lung injury can be protected with anisodamine, and the gomphosis pathway(administer through priming solution and main pulmonary artery) is better. The protective mechanism may be that anisodamine inhibits the activation of NF-κB.

Abstract:Objective:To evaluate the diagnosis of CT perfusion in chronic obstructive pulmonary disease(COPD). Methods:Twenty volunteers and 20 COPD sufferers underwent MSCT perfusion imaging. Cine scan slice with thickness 5 mm/1i were processed by 8-row detector spiral CT. Scan time was 0.5 s per circle, delay of time 5 s, and overall scan time was 45 s. Omnipaque(300 mg/ml) was administered at a rate of 4 ml/s from forearm superficial vein for a total of 60 ml by high pressure injector. The data were transferred to a workstation and analyzed by a CT perfusion 3 software, automatically generating a time-density curve and pseudo-color map. Interest district blood flow, blood volume, mean transit time and permeability surface were measured. Results:Time-density curve of COPD patients was flatter than that of the volunteers, and peak height was obviously lower. The COPD sufferers’ blood flow, blood volume and permeability surface were obviously lower than the volunteers’, which had statistical significance(P < 0.05). Conclusion:MSCT perfusion is helpful to the diagnosis of COPD.

Abstract:Objective:To make a comprasion in surgical stress between laparotomy and transanal pull-throug operation on Hischs-prung-蒺s disease(HD) in children. Methods:Thirty patients were randomly divided into laparotomy group(n=15) and transanal pull-through group(n=15). The blood was collected in the morning before operation, 24 and 72 h postoperation. Plasma levels of IL-6 were determined by ELISA,and C-reaction protein(CRP) levels were measured by immune rate nephelometry. Methods:The statistical difference before and after surgery was analyzed by SPSS software. Results:There were significant difference in plasma levels of IL-6 and CRP before and after surgery(P < 0.05). The transanal pull-through group showed lower level of IL-6 and CRP on 24 and 72 h postoperation than laparotomy group(P < 0.05). Conclusion:Transanal pull-through operation causes less surgical stress than laparotomy on HD.

Abstract:Objective:To analyze the efficacy of local intra-arterial thrombolysis using urokinase in patients with acute ischemic vertebral basilar artery(VBA) stroke. Methods:Ninety-three patients were angiographically examined after VBA stroke. Forty-three patients were treated with local intra-arterial thrombolysis using urokinase,others were control group. The curative effect was compared and the relationship of revascularization 6 h before and after onset with the Glasgow outcome scale(GOS) scores three months later was analyzed. Results:Angiography showed occlusion of VBA in 93 patients. There was significant difference between the therapeutic group and control group(P < 0.01). In thrombolytic group, there were no significant difference between revascularization 6 h before and after onset in clinical prognosis(P > 0.05). Successful revascularization was achieved in 23 patients. Four patients had revascularization and good prognosis with GOS three months later in 6 patients who thrombolysised within 6 h,Sixteen patients had good prognosis with GOS three months later in 19 patients who had successful revascularization among 37 patients who thrombolysised after 6 h. Twelve patients died. Hemorrhage occurred in one patient. Reperfusion injury occurred in 19 patients, and arterial reocclusion occurred in one patient. There was a significant relationship between arterial recanalization rate and clinic prognosis(P < 0.01,r = 0.99). There was low ratio of successful recanalization in VBA with poor clinic prognosis in patients who thrombolysised after 6h. Conclusion:Successful recanalization of VBA occlusive could improve intra-arterial thrombolysis,which is an effective therapy for acute ischemic VBA infarction.There was significant relationship between arterial recanalization rate and clinic prognosis.

Abstract:objective:To establish optimized preparation method of rat osteoblasts secretome sample for two-dimension electroph-oresis(2-DE) and set a foundation for further study of osteoblasts secretome. Methods:By using dialysis, dialysis-trichloracetic acid(TCA)-acetone precipitation and dialysis-acetone precipitation methods respectively, the rat osteoblasts secretome samples were prepared, and then the most optimized preparation method was determined by comparing the 2-DE results. Results: Dialysis-acetone precipitation method was the most optimized method, which lost less proteins, increased its solubility, made 2-DE images more clearly and obtained high resolution. Conclusion:Dialysis-acetone precipitation method is the most optimized preparation method for rat osteoblasts secretome 2-DE sample.

Abstract:Objective:To describe the prevalence rate of metabolic syndrome(MS) in Nanjing district. Methods:1 130 residents from two districts of Nanjing, who had lived for at least five years and more than 20 years old, were enrolled in this survey. Fasting plasma glucose, triglycerides(TG), high density lipoprotein-C(HDL-C) were measured. Results:The crude prevalence of MS were 19.3%, 12.8%, 22.9% in total population, male and female respectively while the standardized rate were 13.1%, 9.2%, 15.2% accordingly. Female and aged people was the population at high risk of MS. The prevalence of hypertension(≥130/85 mmHg),hypertriglyceridemia, abnormal fasting plasma glucose, hypoalphalipoproteinemia-C were 89.9%, 65.6%,29.4%, 71.1% respectively among MS individuals, of which 51.8% incorporated only two metabolic abnormities, 40.4% incorporated three metabolic abnormities and 7.8% had four metabolic abnormities. Conclusion:The prevalence rate of MS in Nanjing was very high. Old and female people were at higher risk. Hypertension was the most frequent metabolic abnormity among MS individuals and the prevalence of other metabolic abnormities were also very high.