Objective To examine the antiplatelet effect of ticagrelor by microfluidic chip and flow cytometry under shear stress in vitro. Methods Microfluidic chip was used to examine the effect of ticagrelor on platelet aggregation at the shear rates of 300/s and 1500/s.We adopted the surface coverage of platelet aggregation to calculate the half inhibition rate of ticagrelor.The inhibitory effect of ticagrelor on ADP-induced platelet aggregation was verified by optical turbidimetry.Microfluidic chip was used to construct an in vitro vascular stenosis model,with which the platelet reactivity under high shear rate was determined.Furthermore,the effect of ticagrelor on the expression of fibrinogen receptor (PAC-1) and P-selectin (CD62P) on platelet membrane activated by high shear rate was analyzed by flow cytometry. Results At the shear rates of 300/s and 1500/s,ticagrelor inhibited platelet aggregation in a concentration-dependent manner,and the inhibition at 300/s was stronger than that at 1500/s (both P<0.001).Ticagrelor at a concentration ≥4 μmol/L almost completely inhibited platelet aggregation.The inhibition of ADP-induced platelet aggregation by ticagrelor was similar to the results under flow conditions and also in a concentration-dependent manner.Ticagrelor inhibited the expression of PAC-1 and CD62P. Conclusion We employed microfluidic chip to analyze platelet aggregation and flow cytometry to detect platelet activation,which can reveal the responses of different patients to ticagrelor.
Humans ; Ticagrelor/pharmacology* ; Platelet Aggregation Inhibitors/pharmacology* ; Flow Cytometry/methods* ; Microfluidics ; Platelet Aggregation

Objective To observe the inhibitory effect of Tirofiban on different shear-induced platelet aggregation, and to provide medication suggestions for the treatment of thrombosis in different hemodynamic environment. Methods Polydimethylsiloxane ( PDMS)-glass microchannel chips were fabricated by soft lithography. The whole blood of healthy volunteers anticoagulated with sodium citrate was collected and incubated with different concentrations of Tirofiban in vitro. The blood flowed through the straight microchannel or channel with 80% narrow for 150 seconds at the speed of 11 μL/ min and 52 μL/ min, respectively. The wall shear stress rates in straight channel at 11 μL/ min and 52 μL/ min were 300 s-1 and 1 500 s-1, respectively. The maximum wall shear rates in the channel with 80% occlusion at 11 μL/ min and 52 μL/ min were 1 600 s-1 and 7 500 s-1, respectively. The adhesion and aggregation images of fluorescent labeled platelets on glass surface were photographed with the microscope, and the fluorescent images were analyzed with Image J. The platelet surface coverage ratio was used as a quantitative index of platelet aggregation behavior, and the IC50 of Tirofiban for platelet inhibition was calculated under different shear rates. Flow cytometry was used to detect the platelet activation index (CD62P, PAC-1) in the whole blood at 52 μL/ min in channel with 80% occlusion. Results Tirofiban inhibited platelet aggregation in a dose-dependent manner, and the inhibitory effect was related to the shear rate. Under the shear rates of 11 μL/ min and 52 μL/ min, the aggregation was almost completely inhibited when the concentration in straight channel reached 100 nmol / L. When the concentration in channels with 80% occlusion reached 1 μmol / L, the aggregation was almost completely inhibited. IC50 values at 11 μL/ min and 52 μL/ min in straight channel were 2. 3 nmol / L and 0. 5 nmol / L, respectively. IC50 values at 11 μL/ min and 52 μL/ min in channels with 80% occlusion were 20. 73 nmol / L and 4. 5 nmol / L. Pathologically high shearforce induced an increase in platelet activation, which could be inhibited by Tirofiban. Conclusions Tirofiban can effectively inhibit shear-induced platelet aggregation, and different concentrations of Tirofiban should be given according to the thrombus formed in different shear force environment in clinic practice

OBJECTIVE: To study the effect of gradient shear stress on platelet aggregation by microfluidic chip Technology. METHODS: Microfluidic chip was used to simulate 80% fixed stenotic microchannel, and the hydrodynamic behavior of the stenotic microchannel model was analyzed by the finite element analysis module of sollidwork software. Microfluidic chip was used to analyze the adhesion and aggregation behavior of platelets in patients with different diseases, and flow cytometry was used to detect expression of the platelet activation marker CD62p. Aspirin, Tirofiban and protocatechuic acid were used to treat the blood, and the adhesion and aggregation of platelets were observed by fluorescence microscope. RESULTS: The gradient fluid shear rate produced by the stenosis model of microfluidic chip could induce platelet aggregation, and the degree of platelet adhesion and aggregation increased with the increase of shear rate within a certain range of shear rate. The effect of platelet aggregation in patients with arterial thrombotic diseases were significantly higher than normal group (P<0.05), and the effect of platelet aggregation in patients with myelodysplastic disease was lower than normal group (P<0.05). CONCLUSION The microfluidic chip analysis technology can accurately analyze and evaluate the platelet adhesion and aggregation effects of various thrombotic diseases unde the environment of the shear rate, and is helpful for auxiliary diagnosis of clinical thrombotic diseases.
Humans ; Microfluidics ; Platelet Adhesiveness ; Platelet Aggregation ; Blood Platelets/metabolism* ; Platelet Aggregation Inhibitors/pharmacology* ; Platelet Activation/physiology* ; Thrombosis

The current review gives a comprehensive overview of the recent development in Chinese medicine (CM) for treating several kinds of acquired nerve deafness and tinnitus, as well as links the traditional principle to well-established pharmacological mechanisms for future research. To date, about 24 herbal species and 40 related ingredients used in CM to treat hearing loss and tinnitus are reported for the treatment of endocochlear potential, endolymph growth, lowering toxic and provocative substance aggregation, inhibiting sensory cell death, and retaining sensory transfer. However, there are a few herbal species that can be used for medicinal purposes. Nevertheless, clinical studies have been hampered by a limited population sample, a deficiency of a suitable control research group, or contradictory results. Enhanced cochlear blood flow, antiinflammatory antioxidant, neuroprotective effects, and anti-apoptotic, as well as multi-target approach on different auditory sections of the inner ear, are all possible benefits of CM medications. There are numerous unknown natural products for aural ailment and tinnitus identified in CM that are expected to be examined in the future utilizing various aural ailment models and processes.
Humans ; Tinnitus/drug therapy* ; Medicine, Chinese Traditional ; Hearing Loss/drug therapy*

OBJECTIVE To study the mitigation effect and its possible mechanism of Astragalus membranaceus polysaccharide on the pulmonary hypertension induced by monocrotaline in rats. METHODS One hundred SD male rats were randomly divided into normal control group ，monocrotaline group ，A. membranaceus polysaccharide low-dose and high-dose groups. In addition to the normal control group, rats in other groups were injected with monocrotaline by single intraperitoneal injection of 60 mg/kg. On days 2 to 28 after administration ，rats in the A. membranaceus polysaccharide low-dose and high-dose groups were intraperitoneally injected with A. membranaceus polysaccharide of 200 mg/kg and 400 mg/kg，respectively，once a day. There were 25 rats in each group，and 15 rats were taken for index detection. The mean pulmonary artery pressure （mPAP）and right heart hypertrophy index （RVHI）were detected ，and morphology changes of pulmonary artery and cardiomyocytes were monitored . mRNA and protein expression of IL- 17 in their lung tissues were detected. RESULTS Compared with normal control group ，mPAP and RVHI of monocrotaline group and A. membranaceus polysaccharide groups were increased significantly （P＜0.01）；mRNA and protein expression of IL- 17 in lung tissues were significantly increased （P＜0.01），and there were obvious pathological changes in pulmonary artery and cardiomyocytes. Compared with monocrotaline group ，mPAP and RVHI were significantly decreased in A. membranaceus polysaccharide groups （P＜0.01），while mRNA and protein expression of IL- 17 in lung tissue were decreased significantly （P＜0.01）；pathological changes in pulmonary artery and cardiomyocytes were improved. Compared with A. membranaceus polysaccharide low-dose group ，above indexes and pathological changes were improved significantly in high-dose group. CONCLUSIONS A. membranaceus polysaccharide can reduce monocrotaline-induced pulmonary hypertension ，improve pulmonary artery structure and myocardial remodeling in rats ， the mechanism of which is presumably related to the down-regulation of IL- 17 expression in lung tissue of rats.

【Objective】 To study the status and conduct effect evaluation of blood donation recruitment of blood services in Chongqing, and explore its influencing factors, so as to provide reference for the regional homogenization of blood services in Chongqing. 【Methods】 19 blood services in Chongqing were investigated by questionnaire in terms of the input in human resources and funds, recruitment methods, document construction and effect evaluation. The statistical analysis was conducted. 【Results】 The average number of blood donors per 1 000 population in 19 blood services in Chongqing was 9.35±3.35. Among the 19 blood services, blood inventory warning occurred in 18, 6 of them reached Level 2 and 1 of them was Level 1. The number of blood donations per 1 000 population in blood banks with no more than 5 recruits or with less than 100 000 yuan/year recruitment fund was significantly lower than that in blood banks with more than 5 recruits or with more than 100 000 yuan/year recruitment fund(P<0.05). SMS and telephone recruitment were most commonly used in blood donation recruitment. Most blood banks have established corresponding system documents, but only one has established the method to evaluate the effect of blood donation recruitment. 【Conclusion】 The number of blood donations per 1 000 population in 19 blood services in Chongqing varies greatly, and the pressure of blood inventory warning is widespread. The input of human resources and financial fund have a certain impact on the number of blood donations per 1000 population, but not the alone factor. The recruitment method is a little bit more on the traditional side, and the blood donation recruitment and efficacy evaluation is in lack of documentary supporting. Regional homogenization should be achieved by integrating the resources of blood services, establishing the document framework of blood donation recruitment and effect evaluation, clarifying the evaluation content and unifying the evaluation standard.

Objective:To explore the mechanism by which SRY-related high mobility group-box 9 (SOX9) promotes the epithelial mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC) A549 cells via the Wnt/β-catenin pathway. Methods: The human NSCLCA549 cell line was divided into three groups: OE-NC group, OE-SOX9 group and OE-SOX9+XAV-939 group. The cells in OESOX9 group were transfected with SOX9 pcDNA plasmid to up-regulate the expression level of SOX9; The cells in OE-SOX9+XAV939 group were transfected with SOX9 pcDNA plasmid while the β-catenin inhibitor XAV-939 (1.0 μmol/L) was added to the medium. qPCR was used to detect SOX9 mRNA levels; CCK-8 was used to examine the proliferation of A549 cells; Wound-healing assay and Transwell chamber assay were used to detect the migration and invasion ofA549 cells, respectively; and WB was used to detect protein expressions of SOX9, β-catenin, E-cadherin, γ-catenin, N-cadherin and vimentin. Results: The mRNA and protein levels of SOX9 in OE-SOX9 group and OE-SOX9+XAV-939 group were significantly higher than those in the OE-NC group after transfection (all P< 0.05), while there was no significant difference between the OE-SOX9 group and the OE-SOX9+XAV-939 group (P>0.05). The proliferation, migration and invasion of cells in OE-SOX9 group were significantly higher than those in OE-NC group; however, those abilities in OE-SOX9+XAV-939 group were significantly lower than those in OE-SOX9 group (all P<0.05). The level of β-catenin protein in OE-SOX9 group was significantly higher than that in the OE-NC group, while the level of β-catenin protein in OE-SOX9+XAV-939 group was lower than that in OE-SOX9 group (all P<0.05). Compared with the OE-NC group, the levels of phenotypic markers of epithelial cells, E-cadherin and γ-catenin, were down-regulated, and the phenotypic markers of mesenchymal cells, N-cadherin and vimentin, were up-regulated in cells of OE-SOX9 group; however, E-cadherin and γ-catenin were higher, and N-cadherin and vimentin were lower in OE-SOX9+XAV-939 group than those in OE-SOX9 group (all P<0.05). Conclusion: SOX9 could promote proliferation, migration and EMT of NSCLCA549 cells by activating the Wnt/β-catenin pathway. ··

Objective: To explore the effect of enteral nutrition on tumor cell proliferation activity in rectal cancer patients with nutritional risk treated with preoperative neoadjuvant therapy. Methods: Sixty-six rectal cancer patients with nutritional risk treated with preoperative neoadjuvant therapy from January 2016 to January 2018 in the Yongchuan Hospital Affiliated to Chongqing Medical University were selected. The patients were divided into experimental group (enteral nutrition combined with neoadjuvant therapy) and control group (simple adjuvant therapy) according to the random digits table method, with 33 cases in each group. The expressions of proliferating cell nuclear antigen (PCNA) and Ki-67 antigen before and after treatment were detected by immunohistochemical method; the albumin and prealbumin before and after treatment were observed, and the nutrition risk screening 2002 (NRS2002) was evaluated. Results: There were no statistical differences in the expressions of PCNA and Ki-67 antigen before treatment between 2 groups (P>0.05); the expressions of PCNA and Ki-67 antigen after treatment in experimental group were significantly lower than those in control group, and there were statistical differences (P<0.05). There were no statistical differences in the NRS2002 score, albumin and prealbumin before treatment between 2 groups (P>0.05); the NRS2002 score after treatment in experimental group was significantly lower than that in control group: (1.58 ± 0.50) scores vs. (3.65 ± 0.72) scores, the albumin and prealbumin after treatment were significantly higher than those in control group: (35.92 ± 2.77) g/L vs. (31.12 ± 1.76) g/L and (204.58 ± 23.86) mg/L vs. (157.46 ± 18.99) mg/L, and there were statistical differences (P<0.01). Conclusions Enteral nutrition can reduce the proliferation activity of tumor cell in rectal cancer patients with nutritional risk treated with preoperative neoadjuvant therapy, and it can improve the nutritional status of patients.

Objective To develop and verify a medical microdevice for analyzing the three-dimensional(3D)migration of tumor cells in extracellular matrix. Methods The mold of the microdevice was made by precision machining,and then the medical microdevice based on polydimethylsiloxane(PDMS)-glass was obtained by PDMS casting,moulding,and bonding.During the analysis,the suspension of tumor cells and matrigel were mixed and then added into the migration channel of microdevice,and the controllable migration of tumor cells in matrigel was induced by establishing chemokine concentration gradient on both sides of the migration channel.Meanwhile,the migration process of tumor cells was recorded with the live cell dynamic imaging device. Results Breast cancer cell line MCF-7 was taken as an example to verify the feasibility of the microdevice to control and dynamically monitor the 3D migration process of tumor cells in vitro.Qualitative analysis of imaging data showed that the migration of MCF-7 cell lines in matrigel was determined by the concentration gradient distribution direction of chemokine and presented as the amoeboid-like migration mode.The proportion and migration velocity of MCF-7 cells could be quantified by the quantitative analysis of cell migration process.The inhibition ability of matrix metalloproteinase inhibitors(Batimastat)and adenosine triphosphatase inhibitors(Blebbistation)on the 3D migration behavior of MCF-7 cells was found to be different.Conclusion This device can be used for in-depth analysis of tumor cell migration and its mechanism and for evaluating the efficacy of anti-metastatic drugs.
Biomedical Research ; instrumentation ; Cell Movement ; Extracellular Matrix ; Humans ; MCF-7 Cells

Objective To explore the inhibitory effect of aspirin and clopidogrel on platelet adhesion and aggregation behaviors under the physiological flow condition using microfluidic chip technology for health volunteers. Methods Peripheral venous blood samples collected from twelve randomly recruited health volunteers were treated with 20 μmol/L acetylsalicylic acid,50 μmol/L 2-methlthioadenosine-5'-monophosphate triethylammonium salt,and their combination,respectively,with untreated blood samples being control group. The blood samples were flowed through a microchannel modified with type I collagen protein at a physiological relevant shear rate of 1000 s for 300 s,while the fluorescent images of platelet aggregations were dynamic captured using a microscope. Based on the images,the platelet coverage rates were calculated and used as quantitative parameters for evaluating platelet adhesion and aggregation behaviors. Results Under a flow condition of 1000 s shear rate,an expected in vivo-like platelet adhesion and aggregation behaviors were observed at the surfaces of collagen proteins for control blood samples. Aspirin alone or clopidogrel alone suppressed platelet adhesion and aggregation at the later period of flow(200-300 s),while the combination of aspirin and clopidogrel reduced the adhesion numbers of platelets at the earlier stage of flow(≤150 s) and compromised the stability of platelet aggregation at the later period of flow(200-300 s). The combination showed synergistic effect in inhibiting platelet aggregation. Furthermore,such inhibitory effect was heterogeneous among 12 volunteers. Conclusion This simple microfluidic technology can offer a new technical platform for analyzing the inhibitory effect of antiplatelet drugs.