Objective To investigate the incidence of the aspirin resistance in secondary prevention of cerebral infarction, and the relationship between the aspirin resistance and the cerebral infarction recurrence or other vascular events during the follow-up periods.Methods Aspirin were taken from the first day of admission in 600 patients with cerebral infarction.The platelet aggregation rate was measured after 7-10 days to screen the patients with aspirin resistance or aspirin sensitivity.All patients were followed up for 6 to 24 months and the cerebral infarction recurrence and other vascular events were recorded.Logistic regression model was used to estimate the risk factors of aspirin resistance, vascular events and prognosis.Results Of 600 patients, 150 (25.0% ) patients were resistant to aspirin and 450 (75.0% ) patients were sensitive to aspirin.The proportion of female and diabetes patients, and the level of low density lipoproteins (LDL) in the aspirin resistance group were higher than those in the aspirin sensitivity group.Diabetes (OR = 2.58, 95% CI 1.37-4.85, P=0.003) and high LDL level (OR = 1.89, 95% CI 1.21-2.93, P = 0.005 ) were independent risk factors of aspirin resistance.The incidence of cerebral infarction recurrence and myocardial infarction and all-cause mortality in the aspirin resistance group were all higher than those in the aspirin sensitivity group.Diabetes ( OR = 2.47, 95% CI 1.36-4.65, P = 0.003 ) , atherothrombosis cerebral infarction (OR = 2.13, 95% CI 1.24-3.95, P = 0.023) and aspirin resistance (OR = 3.86,95% CI 1.79-5.87, P = 0.002) were independent risk factors of vascular events during the following-up period.In the patients with aspirin resistance, the risk of the recurrence of vascular events increased 3.86 times.Conclusions The incidence of aspirin resistance is high in secondary prevention of cerebral infarction.Aspirin resistance is closely correlated with cerebral infarction recurrence and other vascular events.

Objective To research the effect of ulinastatin on T-cytoimmunity in patients with infertility undergoing laparoscopic surgery. Methods Forty patients scheduled for receiving laparoscopic surgery were equally randomized into two groups, ulinastatin group (Group U) and control group (Group C). Ulinastatin was given to patients in the Group U at a dose of 20 × 104 U before anesthetic. No ulinastatin was given to patients in the Group C. Patients′venous blood samples for T-lymphocyte subset (CD3+,CD3+CD4+,CD3+CD8+) and CD3+CD4+/ CD3+CD8+ ratio calculation were collected before the surgery (T0) and at 0 h (T1),1st day (T2),3rd day (T3) after the surgery. Results CD3+ had less difference at T1~3 compared with T0 in the Group C but raised obviously at T2~3 in the Group U. CD3+CD4+ were only raised at T3 compared with T0 but raised obviously at T2~3 in the Group U. CD3+CD8+ were raised obviously at T2~3 compared with T0 in the Group C but had less difference in the Group U. CD3+CD4+/CD3+CD8+ had less difference at T1~3 compared with T0 in the Group C but raised obviously at T3 in the Group U. Conclusion The application of ulinastatin in laparoscopic surgery could significantly produce protective effect on T-cytoimmunity.

Recent studies have confirmed that both CD40 molecule and its ligand CD40L played important roles in various stages of atherosclerosis.The critical cell component of atherosclerosis- endothelial cells,macrophages,and smooth muscle cells on which there are expressions of CD40 and CD40L.The combination of both induces human vascular endothelial cells expressing various active media,participating in the formation of atherosclerosis.However,blocking the CD40-CD40L pathway can prevent atherosclerosis or prevent the plaques from progressing.CD40L may participate in thrombosis and activation of platelet.The soluble CD40L levels increase persistently in patients with acute cerebral infarction and acute coronary syndrome.Some drugs may down-regulate CD40L level.It has provided a new approach for preventing the occurrence of vascular events.

Background Clinical studies indicated that the pathogenesis of most corneal dystrophy is associated with the mutation of the transforming growth factor beta-induced (TGFBI) gene.However,the molecular mechanism of mutated TGFBI gene in corneal dystrophy is unclear. Objective The present study was to investigate the expression of the TGFBI gene in human corneal tissue and cells in vitro.MethodsHuman corneal epithelial cells and keratocytes were cultured and passaged,and donor corneal tissue was obtained for the section preparation.RT-PCR was used to detect the expression of TGFBI mRNA in human corneal tissue and cells.Immunofluorescence was used to test the expression of the TGFBI protein in the human corneal tissue,and immunohistochemistry was used to test the expression of the TGFBI protein in human corneal epithelial cells and corneal stromal cells.ResultsRT-PCR analysis showed that TGFBI mRNA could be detected as a 1274 bp band in human corneal tissue and corneal stromal cells,but no TGFBI mRNA was observed in corneal epithelial cells.Immunofluorescence assay revealed that corneal stromal cells were positive ly expressed for the TGFBI protein,but the corneal epithelial cells did not express the TGFBI protein.Immunohistochemistry indicated that the expression of TGFBI was detected the red fluoressence in the cytoplasm of corneal stromal cells;however,no positive response was found in corneal epithelial cells.ConclusionsThe expression of the TGFBI gene occurs in human corneal stromal cells but not in the corneal epithelial cells.This result might be of helpful for studying the function and role of TGFBI gene in pathogenesis of corneal dystrophy.

Background The human transforming growth factor beta-induced gene (TGFBI) is the first determined pathogenic gene to corneal dystrophy.But the molecular genetic mechanism is completely unknown.The study of concerning role of TGFBI is very important for us understand the physiological function of cornea,and the pathogenesis of corneal dystrophy.Objective The vector of human transforming growth factor beta-induced gene (TGFBI) in eukaryotic expression was constructed and transfected into the human corneal epithelial cells in order to explore its influence on the growth of human corneal epithelial cells.Methods Total RNA was extracted from normal donor cornea tissue and cDNA was obtained by reverse transcription.TGFBI cDNA was synthesized by reverse transcription-PCR and cloned into pCMV-N-HA vector and identified by sequencing with PCR and EcoRV,XhoI double restriction endonuclease.The cells were grouped into recombinant pCMV-N-HA-TGFBI plasmid group,pCMVN-HA plasmid group,non-transfected group and pGFP-C2 transfected group.The recombinant pCMV-N-HA-TGFBI plasmid was transfected to human corneal epithelial cells and identified by observing the expression of enhanced green fluorescence protein(EGFP) in the cells.The TGFBI mRNA and proteins were harvested from the cells for real-time PCR analysis and Western blot assay respectively in 58 hours after transfection.The growth of the transfected cells was assessed by Cell Counting Kit-8.The expressions of matrix metalloproteinase(MMP) and tissue inhibitors of matrix metalloproteinase (TIMP) proteins and their mRNA in transfected cells were detected using SYBR fluorescence realtime PCR analysis and Western blot assay.Results The sequencing result of pCMV-N-HA-TGFBI positive clone plasmid showed that amplified TGFBI eDNA inserted into the vector at the correct sequence.EGFP was expressed in transfected cells in 48 hours after transfer of pGFP-C2 with the transfer efficacy 70％.The expression intensity of TGFBI mRNA was significantly higher in recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group,and TGFBI protein was expressed in recombinant pCMV-N-HA-TGFBI plasmid group.No significant difference was found in the A450value among recombinant pCMV-N-HA-TGFBI plasmid group,pCMV-N-HA plasmid group and non-transfected group ( F=3.34,P＞0.05 ).The mRNA level of MMP1,MMP3in the transfected cells was significant elevated but that of TIMP1 was declined in the recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group (all P ＜ 0.05 ).Meanwhile,the expressions of MMP1,MMP3 and TIMP1 proteins appeared the same tendency( all P＜0.05).Conclusions Eukaryotic expression vector harboring human TGFBI eDNA can be successfully constructed and efficiently overexpressed in human corneal epithelial cells.TGFBI gene is involved in the physical and pathological conditions of human corneal epithelial cells by regulating the activity of MMP1,MMP3 and TIMP1.The results offer a new approach for the study of the role of TGFBI in pathogenesis of corneal transparency.

OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.

RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).

CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.

Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.

Objective To discuss radix hedysari flavonoids mechanism of preventing pulmonary fibrosis.Methods Totally 216 SPF Wistar rats were randomized into normal group, model group, prednisone group, radix hedysari flavonoids high, medium, and low dose groups, and then pulmonary fibrosis model was established by intratracheal dripping of bleomycin. From the second day after modeling, every treatment group received gavage with the corresponding dose of medicine at the planned time for 7, 14, and 28 continuous days. Expressions of MMP-2 and TIMP-1 protein were detected by immunohistochemical method.Results When radix hedysari flavonoids high dose group was at the 7th, 14th, 28th day, and medium dose group was at the 14th day, MMP-2 protein expression was lower than model group, similar to prednisone group. When radix hedysari flavonoids high dose group was at the 14th, 28th day and medium dose group was at 14th day, TIMP-1 protein expression was lower than model group.Conclusion Radix hedysari flavonoids can adjust the expressions of MMP-2 and TIMP-1 protein at different phases, and tend to make the balance of MMPs/TIMPs, which may be an effective mechanism for the inhibition of fibrosis process.

Objective To evaluate the relationship and bias of the Stago-CT and CA1500 automatic coagulation analyzer.Meth-ods The relationship and bias of PT,APTT,INR,FIB,TT,DD examined by the Stago-CT and CA1500 automatic coagulation ana-lyzer by using NCCLS EP9-A2.Results For the six items(PT,APTT,INR,FIB,TT,DD)the r2 were 0.996 9,0.969 1,0.967 7, 0.955 8,0.972 6,0.949 6,respectively,and the bias were 2.9,0.88,5.22,1.16,3.48,20.3.Conclusion The five items (PT, APTT,INR,FIB,TT)at a good relationship(r2 >0.95)by the Stago-CT and CA1500 automatic coagulation analyzer except for the DD(r2 =0.949 6);The bias of the five items(PT,APTT,INR,FIB,TT)were within in the United States of demanding that a third of the clinical laboratory of CLIA 88′bias,except for the DD.

Objective To investigate the expression of apoptotic protease cysteinyl aspartate specific proteinase (caspase-3) in cortex neurons of rats with sepsis.Methods Models of rats with sepsis were established by the cecal ligation and puncture (CLP).Totally 70 cases of 30-day-old male Wistar rats were randomly divided into CLP group (n =50) and control group (n =20).In CLP group,CLP was performed in the rats.Neurobehavioral score was measured in 5 rats at 6,12,24 and 48 h after CLP surgery,respectively.Then,they were killed and their brains were removed.The immunohistochemical staining and Western blot were used to detect the apoptotic protein caspase-3 expression in cortex of rats.Control group did not undergo CLP,and the other treatment was the same as CLP group.Results Neurobehavioral scores at 12,24 and 48 h after CLP surgery were significantly lower than that in the control group(t =3.651,3.773,7.155,all P ＜ 0.05),and the scores were gradually decreased,overall situation of rats was getting worse along with the time.Caspase-3 protein expressed only in trace amounts in rat cerebral cortex in the control group by immunofluorescence analysis,however,its expression was significantly increased at 12 h after CLP surgery.Western blot test showed that caspase-3 protein expression in rat cerebral cortex at 6,12 and 24 h after CLP surgery was significantly higher than those in control group (all P ＜ 0.05).Its expression began to increase at 6 h after CLP surgery,and reached the peak at 12 h,then decreased at 48 h.Conclusion The neurobehavioral scores decreases and the expression of apeptosis protease caspase-3 increases in cortex of rats with sepsis brain injury.