Century Clot在大咯血输血中的指导价值研究

目的 探讨Century Clot在大咯血输血中的指导价值。方法 选取收治60例大咯血患者。随机分为研究组和对照组各30例,除原有治疗方案外,研究组行centuryclot指导输血;对照组行凝血四项及血常规指导输血。比较两组咯血停止时间、治疗后总咯血量及输血总量,比较两组治疗前后的凝血功能(包含TT、PT、APTT以及FIB)及凝血功能监测的准确率。结果 研究组咯血停止时间短于对照组,总咯血量及总输血量低于对照组(P<0.05)。治疗前,两组凝血功能无明显差异(P>0.05);治疗后,两组TT、PT及APTT水平均低于治疗前,且研究组均低于对照组(P<0.05);治疗后,两组FIB均高于治疗前,且研究组高于对照组(P<0.05)。研究组误诊率及漏诊率均低于对照组,但差异无统计学意义(P>0.05),准确率高于对照组(P<0.05)。结论 通过Centuryclot指导大咯血患者输血,不仅可精准止血,还可以有效改善患者凝血功能,减少咯血量及节约血制品。

Promising way to address massive intragastric clotting in patients with acute upper gastrointestinal bleeding:A case report

BACKGROUND Massive intragastric clotting(MIC) makes endoscopic therapy difficult in patients with acute upper gastrointestinal bleeding. Literature data on how to address this problem are limited. Here, we report on a case of massive stomach bleeding with MIC that was successfully treated endoscopically using an overtube of singleballoon enteroscopy.CASE SUMMARY A 62-year-old gentleman with metastatic lung cancer was admitted to the intensive care unit due to tarry stools and hematemesis of 1500 mL of blood during hospitalization. Emergent esophagogastroduodenoscopy revealed massive blood clots and fresh blood in the stomach with evidence of active bleeding.Bleeding sites could not be observed even by changing the patient’s position and aggressive endoscope suction. The MIC was successfully removed using an overtube connected with a suction pipe, which was inserted into the stomach with an overtube of a single-balloon enteroscope. An ultrathin gastroscope was also introduced through the nose into the stomach to guide the suction. A massive blood clot was successfully removed, and an ulcer with oozing bleeding at the inferior lesser curvature of the upper gastric body was revealed, facilitating endoscopic hemostatic therapy.CONCLUSION This technique appears to be a previously unreported method to suction MIC out of the stomach in patients with acute upper gastrointestinal bleeding. This technique could be considered when other methods are not available or if they fail to remove massive blood clots in the stomach.

Intraclot Recombinant Tissue-type Plasminogen Activator Reduces Perihematomal Edema and Mortality in Patients with Spontaneous Intracerebral Hemorrhage

The study aimed to investigate the impact of intraclot recombinant tissue-type plasminogen activator （rt-PA） on perihematomal edema （PHE） development in patients with intracerebral hemorrhage （ICH） treated with minimally invasive surgery （MIS） and the effects of intraclot rt-PA on the 30-day survival. We reviewed the medical records of ICH patients undergoing MIS between October 2011 and July 2013. A volumetric analysis was done to assess the change in PHE and ICH volumes at pre-MIS （T1）, post-MIS （T2） and day 10-16 （T3） following diagnostic computed tomographic scans （To）. Forty-three patients aged 52.8±11.1 years with （n=30） or without rt-PA （n=13） were enrolled from our institutional ICH database. The median rt-PA dose was 1.5 （1） mg, with a maximum dose of 4.0 mg. The ratio of clot evacuation was significantly increased by intraclot rt-PA as compared with controls （77.9%±20.4% vs. 64%±15%; P=0.046）. From TI to T2, reduction in PHE volume was strongly associ- ated with the percentage of clot evacuation （p=0.34; P=-0.027）. In addition, PHE volume was positively correlated with residual ICH volume at the same day （p ranging from 0.39-0.56, P〈0.01）. There was no correlation between the cumulative dose of rt-PA and early （T2） PHE volume （p=0.24; P=0.12） or de- layed （T3） PHE volume （p=0.19; P=0.16）. The 30-day mortality was zero in this cohort. In the selected cohort of ICH patients treated with MIS, intraclot rt-PA accelerated clot removal and had no effects on PHE formation. MIS aspiration and low dose of rt-PA seemed to be feasible to reduce the 30-day mor- tality in patients with severe ICH. A large, randomized study addressing dose titration and long-term outcome is needed.

用Sonoclot分析仪测评体外循环后凝血功能的改变

目的用Sonoclot凝血和血小板功能分析仪观察心脏直视手术体外循环前、后凝血功能的改变。方法选择20例体外循环下心脏手术患者,麻醉前及首次鱼精蛋白中和肝素后10 min取静脉血用Sonoclot分析仪检测全血凝血和血小板功能,并进行激活全血凝固时间(ACT)测定。结果体外循环前后ACT差异无显著性(P=0.27),而使用玻璃珠作为促凝剂的检测试剂盒(gb-ACTKit)术后明显高于术前(P<0.01),凝血速率(CR)术后明显降低(P=0.01),血小板功能(PF)降低但无显著差异(P=0.50)。结论Sonoclot分析仪能够早期检测体外循环后凝血功能的改变,是一种较为敏感的床边检测方法。

Influence of styptic fiber on clotting time in rabbits

Objective To explore the influence of styptic fiber on clotting time in rabbits so as to provide experiment data for its development.Methods Onto 0.1 mL aliquots of citrated anti-coagulant rabbit blood placed in a surfacial plate 25 ul of 0.2 mol·L-1 CaCl2 solution was dropped,and mixed well with glass stirrer;the resulting mixture was immediately capped with a piece of styptic fiber(test product group)or absorptive gelatin sponge(positive control group)of 2 cm diameter.Then,the surficial plate was rinsed with 30ml of purified water at 5,10,20,30,40 and 50 min after capping;the rinsings were allowed to stand for 1 h and were subjected to OD determination at a wavelength of 541 nm.The above procedure was repeated twice,the average value of the twice experiments was taken for evaluation of the hemostatic effect of test product.For negative control group,all procedures except for capping were same as the test product group.The haemostatic effect was judged by percent OD relative to OD at 0 min in negative control group(OD 0 min)(OD 0 min was considered as 100%);if OD value at a time was less than 80% of OD 0 min,it should be designated as primary clotting time(PCT),less than 20% as complete clotting time(CCT).Results The measured PCT was 20min for both negative and positive control groups;CCT was 50,30 and 5 min for negative control,positive control and test product groups,respectively,showing the test styptic fiber had a CCT 8 times shorter than untreated blood,10 times shorter than negative control and 6 times shorter than positive control.Conclusions The test styptic fiber has powerful hemostatic effect.

Blood Flow Through a Catheterized Artery Having a Mild Stenosis at the Wall with a Blood Clot at the Centre

The blood flow through a catheterized artery having a mild stenosis at the wall together with a blood clot at the centre is studied in the current investigation.Stenosis can occur in vessels carrying blood to brain(i.e.,Carotid arteries),Renal arteries that supply blood to kidneys etc.The flow is refined in such vessels by application of catheter.We have used a Newtonian viscous fluid model and also distinct shapes of stenosis,(i.e.,symmetric and non-symmetric shapes)are considered for this study.The entropy generation togetherwith viscous dissipation is also taken into account for a complete description of heat transfer mechanism.Exact solutions are calculated for the problem subject to given“no slip conditions”.The results are discussed graphically.The velocity quickly increases for a non-symmetric stenosis as compared to a symmetric stenosis.When the height of mild stenosis increases and the channel becomes narrow then the velocity increases in the centre but it decreases with the stenosed wall.However,as the height of blood clotσincreases then the velocity of blood flow reduces with the wall having clot but it remains almost same with the stenosed wall.Streamlines are plotted to visualize the flow pattern.The trapping is symmetric for a symmetric stenosis shape but it changes to non-symmetric trapping when we have a non-symmetric shape of stenosis.

Correlation of fibrinogen level and absorbance change in both PT and APTT clotting curves on BCSXP

Objective： To investigate the correlation of fibrinogen level and absorbance change in both PT and APTT clotting curves on BCSXP Analyzer. Methods：A serial of standard fibrinogen and 250 patient plasma samples with different qualities（normal, hemolysis, icterus, and lipemia） were run on BCSXP for assays PT, APTT and Fibrinogen. The absorbance change（DeltaA） from baseline to plateau in clotting curve was retrieved and analyzed on its correlation with the Fibrinogen result. Influence of plasma quality and PT/ APTT result on this correlation was also studied respectively. Results：Both PT-DeltaA and APTT-DeltaA showed good linear regres- sion with fibrinogen level in the sample, with R2 close to 0.90 in both standard and patient samples. Hemolysis（H）, itcterus（I） and lipemia（L） of the sample with valid clotting curves were found to have no significant difference in this correlation from normal（N） sample（R2： 0.83H, 0.92I, 0.81L and 0.79N in PT; 0.89H, 0.95l, 0.91L and 0.89N in APTT） in either PT or APTT curve. PT or APTT result also has little impact on this correlation（0.71 in range 7 - 10 sec, 0.56 inl0 - 20 sec, and 0.70 in 20 sec-; R2 in APTT： 0.88 in 20-30 sec, 0.92 in 30-40 sec, and 0.95 in 40 sec-）. Conclusion：The absorbance change in either PT or APTT clotting curve correlates well with the fibrinogen level in plasma, which is independent of plasma quality PT or APTT results. The absorbance change can be used as an alternative way to roughly estimate fibrinogen level in either PT or APTT clotting curve when the result of clauss-based fibrinogen measurement is not available.

A Case of Adverse Reaction to Booster Dose of COVID-19 Vaccination: Could D-Dimer Elevation Suggest Increased Clotting Risk?

We report the clinical case of a 40-year-old Italian woman, who soon after her booster vaccination with mRNA-1273 after the two previous vaccinations with BNT162b2, developed severe headache, high fever, and Musculo-skeletal pain. She went to the emergency department, where computerized tomography (CT) scans of chest and brain were performed, resulting in both negative for pathologic findings. On the contrary, white blood count was strongly lowered and D-dimer severely elevated. She improved after treatment with enoxaparin and the blood analyses returned in the normal range after ten days. This case supports the hypothesis that COVID-19 vaccines could increase blood clotting in some predisposed subjects. Therefore, we believe that robust and well-designed clinical trials, considering the evaluation of D-dimer levels, should be performed to eliminate any doubts on this issue.

Experimental Results of the Fibrin Clot Use to Accelerate the Regeneration of Damaged Bone in the Rat Lower Jaw

Morphological and radiological methods were used to study regeneration of the damaged bone of rat mandibles after application of plateletenriched fibrin clot. A bone hole was artificially created, and in the natural course of regeneration, the hole was immediately filled with blood and there a blood clot formed. After one week of healing, separate islands of young bone tissue appeared. After two to three weeks, the opening in the mandible was completely replaced by the young bone tissue. When a similar bone hole was filled with autological fibrin clot, the blood clot did not form. But after one week the entire hole was filed with newlyformed fused bone tissue. By the second week after the use of fibrin clot, the bone hole had further healed and bone callus was formed.

Laser induced clot formation in blood treated by EDTA

Patients under systemic (ethylenediaminetetraacetic acid) EDTA are at risk of hemorrhage for any accidental trauma even the superficial ones. This research is a trial to use 980 nm wavelength laser light to provide coagulation for such patients in a local region without collateral thermal damage. Material and Method: 980 nm Diode laser operating in a continuous mode (CW) was incident on human blood treated with EDTA inDurhamtubes. The used parameters were (2.25, 3, 3.75 and 6.75) W output power for variable exposure times (1, 3, 9 and 27) s. Results: A homogenous clot was performed at 4327.84 J/cm2 deposited energy density with the absence of any thermal stress in the blood. Conclusion: 980 nm laser wavelength provides a local photochemical bond break of the anticoagulant enhancing blood coagulation for medically compromised patients.

H∞-Feedback Control of Heparin-Controlled Blood Clotting Network for Cardiac Surgeries

This paper presents a solution methodology for H∞-feedback control design problem of Heparin controlled blood clotting network under the presence of stochastic noise. The formulaic solution procedure to solve nonlinear partial differential equation, the Hamilton-Jacobi-Isaacs equation with Successive Galrkin’s Approximation is sketched and validity is proved. According to Lyapunov’s theory, with solutions of the nonlinear PDEs, robust feedback control is designed. To confirm the performance and robustness of the designed controller, numerical and Monte-Carlo simulation results by Simulink software on MATLAB are provided.

Determination of Rennet Clotting Time by Texture Analysis Method

In this study, texture analysis method was used for the determination of rennet flocculation time (tfloc) and rennet clotting time (tclot) of rennet-induced reconstitued milk samples with different CaCl2 concentrations. The rennet flocculation time (RFT) and rennet clotting time (RCT) were also determined by using the Berridge test and sensory evaluation. The hardness value versus renneting time curves derived from texture analysis gave a good modified exponential relationship for each CaCl2 concentration and the curves were used to calculate flocculation time and clotting time parameters. It was found that the parameters (tfloc and tclot) appeared strongly correlated with RFT and RCT, respectively. Texture analysis was proved as a suitable method to control the rennet-induced coagulation and determine the rennet clotting time. It was also determined that enrichment of milk with CaCl2 leaded to a decrease in flocculation and clotting times and an increase in rate of clotting and gel hardness.

CLOT EXPRESS

CLOT ROYALE是CLOT年度最重头的系列，除了之前看过的一系列联名产品及夏日必备，秋冬的产品也陆续出现．而且陈冠希还亲自上身示范。

Purification and Identification of a Clotting Protein from the Hemolymph of Chinese Shrimp (Fenneropenaeus chinensis)

The clotting protein(CP) plays important and diverse roles in crustaceans,such as coagulation and lipid transportation.A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis(named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography.Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca2+,suggesting that the clotting reaction is catalyzed by a Ca2+-dependent transglutaminase in shrimp hemocytes.The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE.CP exists as disulfide-linked homodimers and oligomers.The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon,Farfantepenaeus paulensis and Litopenaeus vannamei;and similar to that of other decapods.The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry.Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.

Effects of low molecular weight heparin on clot rate and activated clotting time： an in vitro study

Background Due to lack of point-of-care testing, the use of low-molecular-weight heparin （LMWH） therapy in some special patients is restricted. This study was designed to explore the effects of LMWH on clot rate （CR） and activated clotting time （ACT）, and to search for an appropriate method for bedside monitoring of anticoagulant activity of LMWH. Methods Thirty-two healthy volunteers were selected from the staff of Beijing Tongren Hospital. CR and ACT were measured with different reagents （glass beads, diatomite, kaolin and magnetic bar） on blood samples spiked with increasing concentrations of LMWH （dalteparin, 0.2-1.8 IU/ml）. Correlations between concentrations of LMWH and values of CR and ACT were analysed based on the data obtained and regression analysis was performed to establish a regression equation. Results With the increase in doses of dalteparin, CR values reduced gradually. The values of CR of four reagents （glass beads, diatomite, kaolin and magnetic bar） were 20.4-4.5 IU/min, 27.4-6.9 IU/min, 27.5-7.9 IU/min and 7.8-0.1 IU/min respectively and an linear relationship was observed between the CR values and dalteparin concentrations （P〈0.05）. The values of ACT were 173-615 seconds, 130-270 seconds, 123-226 seconds, 337-1411 seconds respectively, which showed a linear regression between the ACT values and dalteparin concentrations （P〈0.01）. Differences in slope of the regression curves of ACT were observed with all the reagents tested （glass beads 248.2 s/IU, diatomite 74.8 s/IU, kaolin 58.2 s/IU and magnetic bar 1112.2 s/IU, P〈0.01）. While the minimum concentration of dalteparin was 0.2 IU/ml, 0.4 IU/ml, 1.4 IU/ml and 0.2 IU/ml separately, the ACT values of the four coagulants （glass beads, diatomite, kaolin and magnetic bar） were beyond the normal limit and showed a noticeable increase respectively （P〈0.01）. Conclusions This study showed that there was an excellent linear relationship between the CR and ACT values and dalteparin concentrations for all the four reagents （glass beads, diatomite, kaolin and magnetic bar） in vitro. The sensitivity of different coagulation reagents to LMWH different. Choosing a suitable reagent, both CR and ACT were possible to be used as a convenient bedside test for LMWH.

Simple procedure for assessing diffuse subarachnoid hemorrhage successfully created using filament perforation method in mice

The murine model of subarachnoid hemorrhage(SAH)is a valuable experimental tool for investigating molecular and cellular mechanisms,and the endovascular filament perforation technique can be used to simulate prominent pathophysiological features observed after human SAH;however,current validation methods for assessing an appropriate SAH model are limited.Here,we introduce a simple procedure for se-lecting a mouse model of diffuse SAH.SAH was induced in 24 mice using a standard filament perforation method.After confirming survival at 24 h,SAH was scored 0-1 based on T2*-weighted images on whole-brain magnetic resonance imaging(MRI)and visual surveillance of the cisterna magna(CM)through the dura mater.The CM-based SAH grading correlated well with a reference parameter defined by extracted brain(r^(2)=0.53,p<0.0001).The receiver operating characteristic curve revealed a sensi-tivity of 85%and a specificity of 91%for detecting diffuse SAH,with a similar area under the curve(0.89±0.06[standard error of the mean])as the MRI-based grading(0.72±0.10,p=0.12).Our data suggest that confirming an SAH clot in the CM is a valuable way to select a clinically relevant diffuse SAH model that can be used in future experimental studies.

小剂量肝素钠与低分子肝素钙对脓毒症治疗作用的对比研究

目的:对比小剂量肝素钠与低分子肝素钙治疗脓毒症的效果。方法:选取2021年10月—2022年10月脓毒症患者100例,随机分为观察组和对照组,每组50例。对比两组患者的治疗效果。结果:对照组弥散性血管内凝血8例(16.0%),观察组弥散性血管内凝血1例(2.0%),观察组临床疗效高于对照组(P<0.05)。随访1个月,观察组病死率低于对照组(P<0.05)。观察组各项凝血指标显著优于对照组(P<0.05)。观察组各项炎性因子水平低于对照组(P<0.05)。两组治疗后急性生理与健康评分(APACHEⅡ)优于治疗前,且观察组优于对照组,差异有统计学意义(P<0.05)。结论:低分子肝素钙相比小剂量肝素钠治疗脓毒症更加可行,可降低并发症发生率,改善短期预后。

益气活血汤联合艾灸治疗四肢骨折术后气虚血瘀证疗效研究

目的:探讨益气活血汤联合艾灸对四肢骨折术后气虚血瘀证患者的影响。方法:选取四肢骨折术后气虚血瘀证患者,随机分为对照组和观察组,每组60例,均连续治疗14 d。对照组接受常规康复治疗,观察组在对照组基础上联合益气活血汤和艾灸治疗。比较两组患者治疗前后的中医证候积分、患肢功能恢复情况、患肢肿胀度、骨代谢状态、凝血功能及并发症发生率。结果:治疗后,观察组患者中医证候评分各项数值低于对照组(均P<0.05)。观察组患者四肢功能恢复理想率高于对照组,观察组活化部分凝血活酶时间、凝血酶原时间、凝血酶时间长于对照组,差异有统计学意义(均P<0.05)。观察组骨碱性磷酸酶、骨钙素、碱性磷酸酶水平高于对照组,而Ⅰ型胶原羧基端肽低于对照组,差异有统计学意义(均P<0.05)。观察组术后相关并发症总发生率低于对照组,差异有统计学意义(P<0.05)。结论:益气活血汤联合艾灸治疗四肢骨折术后气虚血瘀证可有效改善患肢功能恢复状况,加速抗凝血作用,促进术后患者恢复,减少相关并发症发生。

贝莱斯芽孢杆菌YH-1凝乳酶对切达干酪成熟特性及生物活性的影响

为了研究贝莱斯芽孢杆菌YH-1凝乳酶对切达干酪品质的影响,以商品凝乳酶制作的切达干酪(干酪A)为对照组,以70%的商品凝乳酶和30%的YH-1凝乳酶(干酪B),以及YH-1凝乳酶(干酪C)制作的切达干酪为实验组,比较3组干酪在4℃成熟12周过程中成熟特性和生物活性的变化。结果表明,干酪A和干酪B在干酪得率、总蛋白含量和氯化钠含量等指标之间没有显著差异(P>0.05)。干酪C的得率和脂肪含量低于干酪A,但总蛋白含量更高。与干酪A相比,干酪C在成熟过程中水分含量和pH值显著偏低,其中乳酸乳球菌活菌数没有明显变化,但在第12周的pH 4.6可溶性氮含量和12%(体积分数)三氯乙酸可溶性氮含量高于干酪A。干酪C的硬度显著高于干酪A和干酪B,而干酪B中的游离氨基酸含量高于干酪C和干酪A。干酪B和干酪C在成熟过程中所产生的醇类、酮类、酸类、酯类和苯环类化合物的含量高于干酪A。干酪生物活性研究表明,用YH-1凝乳酶制作的切达干酪能提高干酪的抗氧化能力和铁离子螯合能力。因此,YH-1凝乳酶具有部分代替商品凝乳酶生产干酪的潜力。

In vitro and in silico studies on fibrinolytic activity of nattokinase： A clot buster from Bacillus sp.

BACKGROUND： Nattokinase （NK） is a serine protease enzyme of the subtilisin family. It exhibits a strong fibrinolytic activity. The fibrinolytic enzymes from Bacillus sp. have attracted interest as thrombolytic agents because of their efficiency in the fibrinolytic process including plasmin activation. METHODS： In the present study, VIT garden soil was collected and subjected to isolation process in order to screen for the NK production. Screening for NK enzyme was performed by radial caseinolytic assay. The production of NK enzyme was done in two different production medium for comparative studies. The NK enzyme was purified by gel permeation chromatography. The activity of the purified NK was checked by clot lysis and casein digestion assay. To investigate the structural basis of NK and fibrinogen interaction and also to identify the best binding mode, molecular dynamics and docking studies were performed. RESULTS： Based on the morphological and biochemical characterization, the isolate was identified as Bacillus sp. The overall purification fold of NK was about 3 with the specific activity of 664U/mg and 9.9% yield. Homogeneity of the purified enzyme was analyzed and confirmed by the single band obtained in SDS-PAGE. Molecular weight of the purified protease was estimated as 25 kDa. Purified NK enzyme exhibited 97% of effective clot lysis activity. The NK was docked in to the knob region of the fibrinogen at its binding site using Dock server. A total of 26 residues of fibrinogen and 29 residues of NK constitute the interface region. However, 9 residues offibrinogen （THR238, MET264, LYS266, ARG275, THR277, ALA279, ASN308, MET310, and LYS321） and 8 residues ofNK （GLY61, SER63, THR99, PHE189, LEU209, TYR217, ASN218, and MET222） are involved in intact binding. CONCLUSIONS： A significant amount of NK enzyme was obtained from Bacillus sp. The docking analysis revealed that the NK and fibrinogen adopt an extended binding pattern and interacts with the crucial residues to exhibit their activity.