Analysis of the Inhibitory Effect of Tirofiban on Shear-Induced Platelet Aggregation by Microfluidic Technique / 医用生物力学

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

Study on Platelet Adhesion and Aggregation Induced by Gradient Shear Stress Using Microfluidic Chip Technology / 中国实验血液学杂志

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.

Microfluidic Chip and Flow Cytometry for Examination of the Antiplatelet Effect of Ticagrelor / 中国医学科学院学报

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.

Risk factors and countermeasures of depression in acute stroke patients in Wuhan / 公共卫生与预防医学

Objective To explore the risk factors and countermeasures of depression in stroke patients in Wuhan. Methods A stratified cluster sampling method was used to randomly select 628 hospitalized patients at acute stage of stroke in Wuhan from June 2019 to June 2020. According to HAMD score, the patients were divided into normal group (n=381) and post-stroke depression group (n=247). The risk factors of depression in stroke patients were analyzed by Logistic regression using self-made questionnaire, including gender, age, education level, stroke type, residence, patient compliance to treatment, NIHSS score, ADL score and independent walking, and early intervention measures were proposed. Results Univariate analysis showed that education level, family burden, NIHSS score, ADL score and independent walking were related to the occurrence of depression in stroke patients (P<0.05). Logistic multivariate regression analysis showed that, NIHSS score, ADL score and independent walking were correlated with depression in patients at acute stage after stroke (P<0.05). Conclusion the risk of depression in patients with acute cerebral apoplexy is higher, the patients' adherence to treatment, NIHSS score, ADL scores, situation of independent walking is a cause of depression in patients with cerebral apoplexy risk factors, should improve patients' adherence to treatment as soon as possible and daily life ability, improve nerve function, should pay attention to its early intervention.

Identification of Cordyceps cicadae and Tolypocladium dujiaolongae based on ITS sequences and chemical pattern recognition method / 中国中药杂志

Based on ITS sequences, the molecular identification of Cordyceps cicadae and Tolypocladium dujiaolongae was carried out, and high-performance liquid chromatography(HPLC) fingerprint combined with chemical pattern recognition method was established to differentiate C. cicadae from its adulterant T. dujiaolongae. The genomic DNA from 10 batches of C. cicadae and five batches of T. dujiaolongae was extracted, and ITS sequences were amplified by PCR and sequenced. The stable differential sites of these two species were compared and the phylogenetic tree was constructed via MEGA 7.0. HPLC was used to establish the fingerprints of C. cicadae and T. dujiaolongae, and similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminant analysis(PLS-DA) were applied to investigate the chemical pattern recognition. The result showed that the sources of these two species were different, and there were 115 stable differential sites in ITS sequences of C. cicadae and T. dujiao-longae. The phylogenetic tree could distinguish them effectively. HPLC fingerprints of 18 batches of C. cicadae and 5 batches of T. dujiaolongae were established. The results of CA, PCA, and PLS-DA were consistent, which could distinguish them well, indicating that there were great differences in chemical components between C. cicadae and T. dujiaolongae. The results of PLS-DA showed that six components such as uridine, guanosine, adenosine, and N~6-(2-hydroxyethyl) adenosine were the main differential markers of the two species. ITS sequences and HPLC fingerprint combined with the chemical pattern recognition method can serve as the identification and differentiation methods for C. cicadae and T. dujiaolongae.

Efficacy and Safety of PEG-rhG-CSF in HSC Mobilization in 71 Normal Healthy Donors for Allogeneic Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

OBJECTIVE@#To retrospectively analyze the efficacy and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in hematopoietic stem cell mobilization in 71 normal healthy donors for allogeneic hematopoietic stem cell transplantation (allo-HSCT).@*METHODS@#From March 2018 to July 2019, 71 patients received allo-HSCT in The General Hospital of Western Theater Command were enrolled in the study, a single dose of PEG-rhG-CSF was injected subcutaneously at 12 mg to all the stem cell donors. After injection for 4 days, CD34@*RESULTS@#Seventy-one healthy stem cell donors included 39 males and 32 females with a median age of 38 (16-58) years old. The median number of CD34@*CONCLUSION@#For allo-HSCT donor mobilization, PEG-rh-G-CSF is effective, safe, and convenient, providing more options for HSC mobilization.

3D-Morphological Analysis of Inhibitory Effect of Aspirin on Platelet Aggregation / 中国实验血液学杂志

OBJECTIVE@#To develop a new method for evaluating the inhibitory effect of aspirin on platelet by the three-dimensional (3D) morphological parameters.@*METHODS@#The sodium citrate-anticoagulant peripheral blood samples collected from 12 healthy volunteers were divided into 2 groups: group treated with 200 μmol/L acetylsalicylic acid (ASA), and control group. The platelets in the 2 groups were washed and purified. The purified platelets were added into reaction pools modified with type I collagen and induced to activation and aggregation under static condition. The 3D morphology of the formed platelet aggregate was measured by the laser shape microscopic system. Meanwhile, the platelet function was detected by turbidometric light transmittance aggregometry (LTA).@*RESULTS@#This technology could acquire the shape, height and 3D morphology of the platelet aggregates without label, and could quantify their volume parameters. ASA treatment could obviously change the morphology of platelet aggregates. Compared with the parameters of control group, the volume of platelet aggregates in experimental group decreased significantly (t=8.97, P0.05). The receiver-operating characteristic curve(ROC) analysis showed that the platelet aggregate volume as a parameter to identify the ASA inhibition effect had 91.7% sensitivity and 75% specificity when the cut-off value equal to 1395 μm, and its accuracy and sensitivity were both better than those of platelet aggregates rate measured by LTA method.@*CONCLUSION@#The new method developed for evaluating the ASA inhibition of platelet aggregation may provide a complementary strategy for researching and clinically evaluating of ASA anti-platelet aggregation in future.

Dynamical Analysis of the Inhibitory Effect of Aspirin and Clopidogrel on Platelet Adhesion and Aggregation for Healthy People under Physiological Flow Condition by Microfluidic Chip Technology / 中国医学科学院学报

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.

Innate immune cells and liver regeneration / 中国免疫学杂志

Liver regeneration depends on powerful immune system of ownself. TNF-α,IFN-γ,IL-6,IL-12,etc. that secreted by innate immune cells such as macrophages,dendritic cells,NK cells and NKT cells,could act on the hepatocytes and regulate liver regen-eration (LR) through corresponding signaling pathways. This article summarizes the mechanism of different innate immune cells on hep-atocytes,clarifies the recent advances of liver innate immune cellsduring liver regeneration process,lay the foundation for revealing the molecular mechanism of the development of liver regeneration and liver diseases, and for the research and development of new therapeutic methods for liver diseases.

Damage control surgical treatment and nursing care to patients with severe abdominal infection / 现代临床护理

Objective To investigate the application of damage control surgical treatment in the patients with severe abdominal infection and its nursing experience. Methods Toally 10 patients with severe abdominal infection were treated with damage control measures, including damage control surgery, improvement of pathological and physiological status of patients and surgical re-repairing, and the corresponding nursing measures including rapid completion of preoperative preparation, prevention of complications, nursing care to abdominal double cannula drainage, enteral nutrition and disease observation and mobilization the subjective initiative of patients. Results The average hosptitalization time was 21 days (14~29 days). There were 2 cases of anastomotic leakage and 1 case of pulmonary infection who all were cured after 14~29 d. Conclusion Damage control surgery is an effective treatment strategy for patients with severe abdominal infection and the corresponding nursing plays a positive role in improving the treatment effect.

Development and application of sheet for postural change during painless colonoscopy / 医疗卫生装备

Objective To develop a sheet for postural change during painless colonoscopy.Methods The sheet was made of medical canvas whose body had a size of 110 cm (length) × 80 (width) cm.The body had a waistband at its each side,and the waistband had 5 nylon Velcro at each side to tie it to the patient's waist.There were 6 canvas handles at the back of the body,which were in parallel with the waistband for drawing the patient during postural change.A tearing mechanism with Velcro existed at the hip area to facilitate the hip exposure during colonoscopy.Two shoes covers were at right under the body to hold the feet when the patient bent his knees.Results The sheet significantly shortened operation time,increased working efficiency and protected the patient's privacy when compared with manual operation.Conclusion The sheet gains high safety,convenience and efficiency for postural change during painless colonoscopy,and thus is worthy promoting in the hospitals.

Regional cerebral blood flow and cognitive function in patients with obsessive-compulsive disorder / Fluxo sanguineo regional cerebral e funcao cognitiva em pacientes com transtorno obsessivo compulsivo

Objective: To explore the relationship between regional cerebral blood flow (CBF) and cognitive function in obsessive-compulsive disorder (OCD). Method: Single-photon emission computed tomography (SPECT) was performed for 139 OCD patients and 139 controls, and the radioactivity rate (RAR) was calculated. Cognitive function was assessed by the Wisconsin Card Sorting Test (WCST). Results: The RARs of the prefrontal, anterior temporal, and right occipital lobes were higher in patients than controls. For the WCST, correct and classification numbers were significantly lower, and errors and persistent errors were significantly higher in OCD patients. Right prefrontal lobe RAR was negatively correlated with correct numbers, right anterior temporal lobe RAR was positively correlated with errors, and the RARs of the right prefrontal lobe and left thalamus were positively correlated with persistent errors. Conclusion: OCD patients showed higher CBF in the prefrontal and anterior temporal lobes, suggesting that these areas may be related with cognitive impairment. .

Objetivo: Estudar a relação entre a perfusão sanguínea regional cerebral e as funções cognitivas em pacientes com transtorno obsessivo compulsivo (TOC). Método: Foram estudados 139 pacientes com TOC e 139 controles. As imagens do fluxo sanguíneo cerebral foram feitas por single photon emission computed tomography e o radioactivity rate (RAR) foi calculado usando como referência a radioatividade do córtex cerebelar. As funções cognitivas foram avaliadas através do Wisconsin Card Sorting Test (WCST). Resultados: O RAR do lobo pré-frontal, do lobo temporal anterior e do lobo occipital direito foram mais elevados nos pacientes do que nos controles (P<0.05). Quanto ao WCST, os escores de números corretos (P<0.01) e de classificação (P<0.05) foram menores no grupo TOC do que nos controles; entretanto, os escores de erros e de erros persistentes nos controles foram mais elevados do que nos pacientes (P<0.01). O RAR do lobo pré-frontal direito estava relacionado de modo negativo com os números corretos; o RAR do lobo temporal anterior apresentava correlação positiva com os erros; o RAR do lobo pré-frontal direito e do tálamo esquerdo apresentavam correlação positiva com erros persistentes. Conclusão: Pacientes com TOC apresentaram perfusão sanguínea cerebral mais intensa no lobo pré-frontal e no lobo temporal anterior. O lobo pré-frontal direito, o lobo temporal e o tálamo esquerdo podem estar relacionados com alterações cognitivas. .

1H-NMR based metabonomic approach to evaluate detoxification effect of vinegar-processed Euphorbia kansui / 中国中药杂志

Euphorbia kansui (EK) is a toxic herbal drug, and often used after vinegar-processing to reduce its toxicity. In present study, a 1H-NMR based metabonomic approach was used to evaluate the detoxification effect of vinegar-processed EK. The water extracts of EK and VEK were administered orally to male SD rats at doses of 9 g x kg(-1) x d(-1) for 1 week, respectively, and one more week observation was further conducted. The control group was orally given with saline. Histopathological studies of liver samples on the 8th and 15th day were conducted, and the metabolites of rat urine and liver were analysed by 1H-NMR. Histopathological studies of liver samples from EK and VEK treated rats showed no negative impacts. In metabonomic analyses of urines, changes of metabolites indicated liver damages, kidney lesions and imbalance of gut microbes in the second week. VEK-treated rats showed a quite lower toxicity compared with EK-treated ones. The present study revealed that the metabonomic approach might be helpful for the evaluation of toxicity of EK and detoxic effect of VEK.

Observation on therapeutic effect of the third lumbar transverse process syndrome treated with acupotomy and blood pricking therapy / 中国针灸

<p><b>OBJECTIVE</b>To explore the better treatment for the third lumbar transverse process syndrome.</p><p><b>METHODS</b>Ninety cases were randomly divided into an acupotomy group, a blood pricking group and a combined therapy group, thirty cases in each group. In acupotomy group, three points such as the third lumbar transverse process point were released with acupotomy, and then Tuina was applied. In blood pricking group, blood pricking therapy was applied at Qihaishu (BL 24) or Weizhong (BL 40). In combined therapy group, both of releasing with acupotomy and blood pricking were applied. The therapeutic effects of three groups were evaluated by of simplified Chinese version of the Oswestry Disability Index (SCODI) and Visual Analogue Scale (VAS) before, immediately after treatment and 1 week after treatment.</p><p><b>RESULTS</b>After treatment, compared with scores sofore treatment, VAS scores were all reduced (both P < 0.05), and it in combined therapy group was lower than those in other groups (both P < 0.05); SCODI scores showed no significant differences in groups (both P > 0.05) except in combined therapy group (P<0. 05). One week after treatment, compared with scores immediately after treatment, SCODI scores and VAS scores were reduced in each group (all P < 0.05), and the scores in combined therapy group were lower than those in other two groups (both P < 0.05).</p><p><b>CONCLUSION</b>Releasing with acupotomy and blood pricking are the effective therapies for the third lumbar transverse process syndrome, can remarkably relieve the pain and improve movement function of lumbus, and the therapeutic effect is more obvious.</p>

The surface-electromyographic characteristics of the anterior and posterior thigh muscles in patients recovering from cerebral infarction / 中华物理医学与康复杂志

Objective To assess the contraction and coordination changes in the anterior and posterior thigh muscles of patients recovering from cerebral infarction,and to provide objective references for targeted rehabilitation programs. Methods Eighteen cerebral infarction patients with mild hemiparesis (the patient group) and eighteen age- and sex-matched healthy volunteers (the healthy group) were investigated. The surface-electromyographic (sEMG) signals of their vastus medialis,rectus femoris,vastus lateralis,biceps femoris,semitendinosus and semimembranosus were recorded during knee joint flexion and extension in the prone position.The sEMG signals from both legs were recorded for the patient group,but only from the left leg in the healthy group.The standardized root mean square (stRMS) signals and the co-contraction ratios (CRs) were compared and analysed, Results The stRMSs of the rectus femoris,vastus lateralis and vastus medialis on the patients' affected side during knee extension were significantly higher than those in the healthy group at baseline and follow-up.The stRMSs of the biceps femoris and semitendinosus-semimembranous on the patients' unaffected side were significantly higher than those of the affected side and the healthy group during knee flexion at baseline.The CRs during knee flexion on the patientsˊ affected side at baseline and follow-up were significantly higher than those of the unaffected side at baseline. Conclusions After cerebral infarction,the functioning of both the anterior and posterior thigh muscles on the affected side are impaired.The rectus femoris are the most severely impaired knee extensors,and the biceps femoris,semitendinosus and semimembranous are impaired equally in knee flexion.The thigh flexors and extensors lose their normal antagonist-agonist contraction modes.The functions of both the anterior and posterior thigh muscles should be emphasized during rehabilitation to improve abnormal contraction.

Effect of rocuronium on entropy to endotracheal intubation during propofol anesthesia induction / 中国医师进修杂志

Objective To evaluate the effect of rocuronium on entropy to endotracheal intubation during anesthesia induction with propofol. Methods Forty patients anesthetized induction with propofol using a target-controlled infusion were randomly divided into two groups: rocuronium group (R group, 20 cases) received 0.6 mg/kg rocuronium or saline group (S group, 20 cases) received saline. 2-3 min later, endotracheal intubation was performed. Response entropy(RE) and state entropy(SE) were recorded during baseline(Ta), at steady state(Tb), 2 min after rocuro nium or saline administration (Tc) and 0, 1, 2 and 3 min after endotracheal intubation (T0, T1, T2, T3). Results At T2, the RE-SE was higher in S group than that in R group. Endotracheal intubation induced increasing in RE and SE. Comparing T2 and T0 values in R group and S group, SE increased from 42 ± 7 to 50 ± 8 and 43 ± 13 to 55 ± 12, and RE increased from 45 ± 6 to 54 ± 9 and 48 ± 16 to 66 ± 15, respectively. At T0, RE and RE-SE were higher in S group. Conclusion Rocuronium affects RE-SE and RE and RE-SE responses to endotracheal intubation and may confound interpretation of entropy monitoring.

Expression and purification of an adhesive protein of rabbit Pasteurella multocida C51-3 and detection of its antigenicity / 生物工程学报

The cp36 gene encoding an adhesive protein was amplified by PCR from genomic DNA of rabbit P. multocida C51-3 strain, and cloned into the pMD18-T vector and then sequenced. The mature adhesive protein without a signal peptide of cpm36 gene was amplified by PCR from the recombinant plasmid pMD18-cp36, then cloned into the prokaryotic expression vector pQE30 to provide a recombinant plasmid pQE30-cpm36. The recombinant protein of CPM36 was produced in Escherichia coli M15 harboring the recombinant plasmid pQE30-cpm36 by IPTG induction, and the recombinant protein purified by the affinity chromatography with Ni(2+)-NTA resin. The sequence analyses showed that the ORF of cp36 gene was 1032 bp in length, and DNA homology of the cp36 genes between the C51-3 strain and the previously reported different serotype strains of P. multocida in GenBank was 76.9 to 100%. The SDS-PAGE analyses revealed a single fusion protein band with a molecular weight of 37 kD, and the Western blotting analysis demonstrated that the recombinant protein CPM36 and native 36 kD protein of C51-3 were recognized specifically by an antiserum against the recombinant protein, suggesting that the recombinant protein is an antigenic protein.