Mechanisms of leucocyte recruitment to the inflamed large intestine: redundancy in integrin and addressin usage

The caecal-dwelling nematode Trichuris muris provides a natural model of human whipworm infection. Resistance to T. muris is dependent on a host Th2 response, and CD4+Th2 cells migrate to the gut-associated lymphoid tissue (GALT) to elicit parasite expulsion. Thus, CD4+T cells infiltrate the caecal lamina propria during infection, along with other leucocyte subsets that are not critical for parasite expulsion, such as eosinophils. Trafficking of leucocytes to the GALT has been shown to be dependent on the alpha4beta7/MAdCAM-1 integrin-addressin interaction. However, where inflammation is present, such as during T. muris infection, redundant mechanisms of leucocyte recruitment may also occur in addition to traditional gut-homing interactions. We utilized an anti-integrin/addressin antibody treatment regime to investigate this redundancy in resistant, T. muris-infected C57BL/6 mice. Where only the alpha4beta7/MAdCAM-1 interaction was blocked, mice remained resistant to T. muris infection, making a Th2 response and both CD4+T cells and eosinophils infiltrated the site of infection. However, in the absence of available alpha4beta7 and alpha4beta1, mice became chronically infected with T. muris and mounted a more Th1-biased immune response. Interestingly, CD4+T cells, but not eosinophils, were able to infiltrate the caecum, showing different levels of redundancy between leucocyte subsets during infection.     

Degradation in vivo of articular cartilage in rheumatoid arthritis by leucocyte elastase from polymorphonuclear leucocytes

Summary Using a specific substrate, no leucocyte elastase activity could be detected in 55 synovial fluids, including 29 from patients with rheumatoid arthritis (RA). However, a high percentage of samples contained phagocytic inclusions of elastase, ₁-proteinase inhibitor (₁-PI) and ₂-macroglobulin (₂-MG) in both the polymorphonuclear (PMN) and mononuclear phagocytes. Immunofluorescence and indirect peroxidase-antiperoxidase staining of articular cartilage (ACA) from 52% of 21 patients with RA and one with juvenile RA (JRA) showed presence of elastase in the superficial layer of microscopically intact but proteoglycan depleted pannus-free ACA. In histologically altered pannus-free RA-ACA superficial elastase deposits were found in 24% of the cases. Adjacent ACA sections contained IgG, C 3, ₁-PI and rarely ₂-MG. RA-ACA below or surrounded by pannus showed close contact with intact and decaying PMN in 62% and 48% of the cases, respectively. ACA specimens from patients with degenerative disease and systemic lupus were negative. These findings strongly suggest that PMN leucocyte elastase is operative in the degradation of RA-ACA and JRA-ACA, and that this activity is largely dependent upon the presence of entrapped immune complexes in such cartilage.     

Analysis of leucocyte populations with the Coulter S-Plus STKR as a screening tool for haematological abnormalities

Summary In two institutions at Rome and Florence we evaluated the clinical sensitivity of two Coulter STKR systems using the NCCLS standard H20-T for leucocyte differential count in a patient population with high prevalence of haematologic abnormalities. Reference ranges of normal leucocytes were obtained on 278 adult subjects. On a population of 455 patient specimens, 200 specimens (44%) were flagged by the STKR because of a distributional abnormality, and 122 (27%) because of a morphological abnormality. Percentage of subtotal agreements between the STKR and the reference manual differential count was 85.4%, with 67.5% full and 20.9% partial agreements. Eight specimens that showed a morphological abnormality with the reference manual differential count were classified as normal by the STKR, with a false normal rate of 6.6%. Analysis of the STKR performance for morphological abnormalities showed acceptable sensitivity (82.0%) and rather low specificity (71.5%), low predictive value of positive results (51.3), high predictive value of negative results (91.5%) and efficiency of 74.3%. The main problems of the STKR differential count were a high rate of false monocyte count, and the misidentification of eosinophilias and low-concentration abnormal cells.     

Quantification of leucocyte elastase and cathepsin G in plasma by a simple method: effect of elastase in plasma levels of D-dimer and thrombomodulin

The purpose of this study was to determine levels of leucocyte elastase and cathepsin G in the plasma of patients in various pathological states, in which plasma increases or decreases in coagulation and fibrinolytic factors were seen. Simple methods were developed to measure the leucocyte proteinases and the results were correlated with conventional assays of coagulation and fibrinolytic factors. The total number of patients and total number of plasma samples examined were 340 and 1292, respectively. No correlation was observed between the plasma levels of elastase and cathepsin G, and plasminogen, fibrinogen and leucocyte counts. There was a weak overall correlation, however, between the leucocyte proteinases and each of the four parameters: D-dimer. thrombomodulin, antithrombin III and platelet count. There was a strong correlation between leucocyte proteinases and D-dimer and thrombomodulin in those patients with plasminogen levels within the normal range. Increased D-dimer levels, as well as plasmin, may suggest that elevated leucocyte proteinases contribute to elevated fibrinolytic mechanisms in these instances.     

溶质清除指数量化血液透析充分性的研究

目的寻找一个充分的透析剂量,以提高透析患者生存质量。方法对２０例稳定血液透析患者进行了溶质清除指数的临床研究。检测患者透析前、结束和透析后１小时血尿素氮（ＢＵＮ）水平,用尿素动力学二室模型方法计算出尿素生成量（Ｇ）、蛋白分解代谢率（ＰＣＲ）、溶质排除量（Ｒ）和溶质清除指数（ＳＲＩ）。结果透析后１小时尿素反跳率（１８５０±３３５）％,Ｒ（１３８２±５４８）ｇ,Ｇ（４８５±１３９）ｇ,ＰＣＲ（０９４±０２９）ｇ·ｋｇ－１·ｄ－１,ＳＲＩ（７１３３±６８０）％。结论提示ＳＲＩ是量化透析剂量的一个较好的指标。     

Endoscopic removal of foreign bodies:A retrospective study in Japan

BACKGROUND The ingestion of foreign bodies(FBs) and food bolus impaction(FBI) in the digestive tract are commonly encountered clinical problems.Methods to handle such problems continue to evolve offering advantages,such as the avoidance of surgery,reduced cost,improved visualization,reduced morbidity,and high removal success rate.However,to date,no studies have evaluated the endoscopic management of FBs in Japan.AIM To elucidate level of safety and efficacy in the endoscopic management of FBs and FBI.METHODS A total of 215 procedures were performed at Keio University Hospital between November 2007 and August 2018.Data were collected from medical charts,and endoscopic details were collected from an endoscopic reporting system.Procedures performed with a flexible gastrointestinal endoscope were only taken into account.Patients who underwent a technique involving FB or FBI from the digestive tract were only included.Data on patient sex,patient age,outpatient,inpatient,FB type,FB location,procedure time,procedure type,removal device type,success,and technical complications were reviewed and analyzed retrospectively.RESULTS Among the 215 procedures,136(63.3%) were performed in old adults(≥ 60 years),180(83.7%) procedures were performed in outpatients.The most common type of FBs were press-through-pack(PTP) medications [72(33.5%) cases],FBI[47(21.9%)],Anisakis parasite(AP) [41(19.1%) cases].Most FBs were located in the esophagus [130(60.5%) cases] followed by the stomach [68(31.6%) cases].AP was commonly found in the stomach [39(57.4%) cases],and it was removed using biopsy forceps in 97.5% of the cases.The most common FBs according to anatomical location were PTP medications(40%) and dental prostheses(DP)(40%) in the laryngopharynx,PTP(48.5%) in the esophagus,AP(57.4%) in the stomach,DP(37.5%) in the small intestine and video capsule endoscopy device(75%) in the colon.A transparent cap with grasping forceps was the most commonly used device [82(38.1%) cases].The success rate of the procedure was100%,and complication were observed in only one case(0.5%).CONCLUSION Endoscopic management of FBs and FBI in our Hospital is extremely safe and effective.     

A new approach to the nipple in Kock's reservoir ileostomy using mersilene® mesh

Disinvagination of the nipple in reservoir ileostomy reached an incidence of 33 per cent. The research work we carried out in dogs enabled us to find a solution to this pitfall by using Mersilene^® mesh for reinforcement of the nipple. The good results obtained encourage us to perform this procedure in human beings as well.     

Cytokine removal on extracorporeal life support for treatment of acute endotoxemia: A randomized controlled animal study

Background

We prospectively evaluated the effectiveness of resin adsorption incorporated in an extracorporeal life support (ELS) circuit in an animal model of sepsis for removal of cytokines and prevention of hemodynamic deterioration during the treatment of septic shock.

Methods

Twelve female landrace pigs were randomly assigned to two groups, a study group(n = 6), treated with high-flow resin adsorption (300 mL/min) and ELS, and a control group (n = 6), treated only with ELS. Septic shock was induced by intravenous 0.02 μg/kg/min infusion was of Escherichia coli lipopolysaccharide (LPS). Measurements were carried out in the study group at baseline, at the end of LPS injection(t₀) at 30(t₁), 60(t₂), 90(t₃) and 120 min (t₄) and 60 min after stopping resin-adsorption (t₅). In the control group measurements were performed at baseline (t₀), t₁ and only t₂, as no control animal survived beyond this latter experimental timepoint.

Results

The final population consisted of 9 animals, five in the study group and 4 in the control group. Plasma values of both tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) were reduced during resin-adsorption (t₁–t₄) while these mediators increased in controls undergoing ELS only. With a clearance of TNF-α of 15,233 pg/min and IL-6 of 10,233 pg/min, 79.2% of TNF-α and 95.3% of IL-6 produced were adsorbed. Systemic vascular resistance decreased significantly in both groups at t₀. While it further was reduced during the control experiments at t₁ and t₂, it returned to normal in the study animals. Cardiac output increased at t₀, t₁ and t₂ in the control experiments. In contrast, in study animals after a peak at t₀, it returned to the baseline value and did not vary thereafter.

Conclusions

Combined resin-adsorption and ELS improved hemodynamics resulting from effective removal of inflammatory mediators in a pig model of septic shock.     

Blood pressure and TNF-α act synergistically to increase leucocyte CD11b adhesion molecule expression in the BELFAST study: implications for better blood pressure control in ageing

Hypertension, a key risk factor for stroke, cardiovascular disease and dementia, is associated with chronic vascular inflammation, and although poorly understood, putative mechanisms include pro-inflammatory responses induced by mechanical stretching, with cytokine release and associated up-regulated expression of adhesion molecules. Because blood pressure increases with age, we measured baseline and tumour necrosis alpha (TNF-α)-stimulated CD11b/CD18 adhesion molecule expression on leucocytes to assess any association between the two. In 38 subjects (mean age 85 years), consecutively enrolled from Belfast Elderly Longitudinal Free-Living Aging Study (BELFAST), baseline and TNF-α-stimulated CD11b/CD18 expression on separated monocytes and neutrophils increased with systolic blood pressure >120 mmHg (p = 0.05) and for lymphocytes, with diastolic blood pressure >80 mmHg (p < 0.05).These findings show increased potential stickiness of intravascular cells with increasing blood pressure which is accentuated by TNF-α, and suggest mechanistic reasons why better hypertension control is important.     

Effect of clot removal on cerebrovascular contraction after subarachnoid hemorrhage in the monkey: Pharmacological study

Summary Clot removal at early surgery has been reported to be clinically effective for the prevention of cerebral vasospasm following subarachnoid hemorrhage (SAH) due to rupture of an intracranial aneurysm. We examined the most efficacious timing of mechanical clot removal on pharmacological responses in a monkey SAH model. Cynomolgus monkeys (Macaca fascicularis) were randomized into five groups: sham-operated, clot removal in which the clot was removed 48, 72, or 96h after SAH, and clot groups. An autologous blood clot was placed around the bilateral major cerebral arteries after craniectomy to mimic the hemorrhage. Seven days after the SAH, proximal and successively distal parts of the middle cerebral arteries were cut into rings for isometric tension measurement. The contractile responses to potassium chloride, 5-hydroxytryptamine, norepinephrine, adenosine triphosphate, prostaglandin F₂, and hemoglobin were greater in the proximal parts than in the distal parts in each group. Compared with the sham-operated group, the responses of the clot-removal and clot groups to the drugs were progressively attenuated. The maximum responses to 5-hydroxytryptamine in the proximal parts and to adenosine triphosphate in the distal parts started to decrease, significantly, in the clot-removal group 48h after SAH, while most of the responses to the other agonists began to decrease in the clot-removal groups later than 72h after SAH. These results suggest that the attenuation of cerebrovascular contractile responses 7 days after SAH is pharmacologically inevitable, even if the clot is removed as early as 48h after the SAH. Clot removal may thus be recommended within 48h after SAH to ameliorate the severity of cerebral vasospasm following SAH.     

Evacuation pouchography in the evaluation of ileoanal reservoir function

Thirty four patients with restorative proctocolectomy and ileoanal pelvic reservoirs were studied by “evacuation pouchography” to determine why only some patients could evacuate spontaneously. The 50 per cent who were able had a significantly shorter distal segment (P<0.02) of mean length (8±3 cm) that filled on straining, compared with those who had to use a catheter to empty the pouch, in whom the distal segment was longer (mean 11±4 cm) and often failed to fill on straining. The longer the distal segment, the more likely it was to be angulated, causing difficulty in passage of the catheter with possible ulceration and stenosis from minor trauma. Stricturing in the distal segment was associated with minor nocturnal leakage. A short distal segment is recommended to allow spontaneous evacuation and avoid the risks of repeated catheterization. This work was performed at St. Mark's Hospital.     

A method of test substance removal in agar colony assays using glass capillaries

Summary A technique has been developed to remove test substances, after defined incubation periods, from clonogenic in vitro assays using agar-containing glass capillaries. Following removal from the capillaries, the entire agar gels were washed in petri dishes and redrawn into new capillaries. Using 8 radioactive biochemicals of molecular masses ranging from 150 to 1300 dalton the kinetics of diffusion between 1 and 20 min were determined. Using a wash solution-to-assay volume ratio of 20:1, a single washing for 10 min yielded between 90% and 99% removal by diffusion of test substances. By incorporating myelopoietic stem cells it was demonstrated that the cells to be assayed can be quantitatively transferred, without loss or stress, out of and back into capillaries. Thus the reversibility of test substance action can be examined under defined conditions avoiding technical problems of previous methods.     

CD146在血管炎患者外周血白细胞表达的意义初探

目的探讨血管炎患者外周血白细胞CD146表达与临床活动性间的关系。方法流式细胞术检测39例活动期系统性血管炎患者[显微镜下多血管炎（MPA）13例,韦格纳肉芽肿（WG）9例,变应性肉芽肿性血管炎（CSS）2例,大动脉炎（TA）9例,白塞病（BD）4例,结节性多动脉炎（PAN）2例]及24例系统性红斑狼疮（SLE）患者外周血白细胞CD146表达,其中18例（MPA5例,WG4例,CSS2例,SLE4例,PAN2例,TA1例）患者经糖皮质激素和环磷酰胺治疗后于病情好转时再次检测。结果（1）与健康者相比,血管炎患者活动期中性粒细胞、淋巴细胞CD146表达增多,尤以中性粒细胞最多,差异均有统计学意义（P〈0．05）。（2）中性粒细胞CD146表达与淋巴细胞、单核细胞CD146表达相关（r值分别为0．66、0．853,P=0．000）,与病程、年龄、血沉、C反应蛋白、抗中性粒细胞胞浆抗体（ANCA）、PR3-ANCA、MPO-ANCA、血肌酐、伯明翰血管炎活动指数（BVAS）、系统性红斑狼疮疾病活动指数（SLEDAI）等无明显相关（r值分别为-0．108、-0．059、-0．073、-0．103、0．012、-0．5、-0．232、0．001、-0．08、0．089,P〉0．5）。（3）18例患者经治疗后好转期中性粒细胞、淋巴细胞CD146表达多数呈逐渐减少的趋势（P〈0．05）。结论CD 146在血管炎患者活动期外周血白细胞尤其是中性粒细胞中表达明显升高,随着糖皮质激素和免疫抑制剂治疗病情好转后呈下降或转阴趋势,其在血管炎发病机制中的意义有待深入研究。     

Endoscopic removal of a tablespoon lodged within the duodenum

Here we report the case of a 34-year-old man who underwent endoscopic removal of a tablespoon from the stomach that was lodged within the duodenum.Removal required the use of a two-channel upper endoscope and polypectomy snares.Using the doublesnare technique, the spoon was grasped at the proximal and distal parts of the handle.The doublesnare was first pulled unsuccessfully and then pulled with simultaneous manual abdominal compression of the bulbus from the body surface.Compression was gently applied towards the stomach.As a result, the head of the spoon prolapsed from the bulbus, and was easily retracted from the stomach without any complications.In cases of foreign body lodging within the duodenum, the manual abdominal compression technique may help clinicians pull out the object and avoid surgery.The usefulness of manual compression is dependent on the foreign body's sharpness and the location.     

脑室出血Ommaya储液囊外引流41例临床分析

目的探讨脑室外引流结合Ommaya储液囊置入治疗脑室内出血的临床疗效。方法对脑室内出血患者41例行一侧脑室外引流术,对侧脑室置入引流管接Ommaya储液囊埋植于头皮下经皮穿刺外引流。结果 41例患者中,按GOS预后标准评价疗效,预后良好24例,中残8例,重残4例,植物生存2例,死亡3例,无一例发生颅内感染。结论脑室外引流结合Ommaya储液囊植入引流术能有效降低颅内感染和脑积水的发生率,提高了治疗效果。     

Stanford A型主动脉夹层手术中的体外循环管理

目的:总结我院165例Stanford A型主动脉夹层手术体外循环管理经验。方法:收集165例Stan-ford A型主动脉夹层患者,实施Wheat、David、Bentall及全弓置换和支架象鼻术等术式。根据术式分别采用常规中低温体外循环,深低温停循环(DHCA)加选择性顺行性脑灌注(SACP)等体外循环灌注方法。其中,主动脉全弓置换术均行右锁骨下动脉与右心房插管建立体外循环,在DHCA加SACP下完成主动脉远端支架植入及弓部血管吻合,期间脑灌注流量5～10ml.kg-1.min-1。术中采用单泵双管及单侧选择性脑灌注、α+pH稳态和高氧血气管理及超滤等技术。结果:体外循环转流时间(197.3±28.3)min、深低温停循环时间(25.3±3.8)min、SACP时间(45.2±7.7)min、心肌阻断时间(86.1±10.8)min。重症监护室时间72～516(181.31)h。院内死亡17例(死亡率11.25%),余134例均痊愈出院,出院时心功能NYHAⅠ～Ⅱ级。结论:正确选择体外循环方法及良好的体外循环管理是Stanford A型主动脉夹层手术成功的保障。主动脉全弓替换术中采用单泵双管及单侧选择性脑灌注、α+pH稳态和高氧血气管理及超滤等技术切实可行,临床预后满意。     

核素心肌灌注显像评价FH纯合子患者心肌血供异常临床意义初步研究

目的:利用核素心肌灌注显像(MPI)研究家族性高胆固醇血症(FH)纯合子患者病情进展对心肌血供的影响及临床意义。方法:以经染色体检测确诊并来我院检查的28例FH纯合子患者为研究对象,均行药物负荷/静息MPI,根据心肌血供异常与否,将患者分成两组,并对两组患者的基本资料、血脂水平及心肌血流灌注情况进行对比分析,其中8例患者随诊1~2年。结果:28例FH患者,男性17例,女性11例,平均年龄12.12岁,TC(16.00±2.92)mmol/L,LDLC(13.82±2.73)mmol/L;MPI示15例阴性(男/女=11/4例),13例阳性(男/女=6/7例)。负荷试验中6例出现心电图ST段下移性改变。MPI显示:13例MPI阳性患者共计221个心肌节段中,29个节段出现心肌缺血(13.1%);其中左前降支、回旋支、右冠状动脉供血区分别有21个节段(21/29,72.4%)、5个节段(5/29,17.2%)、3个节段(3/29,10.3%)。MPI阳性与阴性两组中,患者的年龄(P=0.621)、身高(P=0.219)、体质量(P=0.847)、体质量指数(P=0.155)、治疗时间(P=0.189)、性别(P=0.246)、心电图ST段有无下移(P=0.372)、TC(P=0.088)、LDL-C(P=0.082)、TG(P=0.062)、HDL-C(P=0.652)均差异无统计学意义。8例随诊患者复查MPI结果提示:4例原无心肌缺血现仍为阴性;4例原有心肌缺血者,3例心肌缺血范围和程度均无明显变化,1例因心肌缺血程度加重导致心力衰竭后死亡。结论:FH纯合子患者心肌缺血好发于左前降支供血区,且患者的年龄、身高、体质量、治疗时间、血脂等因素在MPI阳性组与阴性组中的差异无统计学意义;最好早期对FH纯合子患者行MPI以评估心血管病危险度分层、患者治疗效果和预后。     

The bone integration effects of platelet-rich fibrin by removal torque of titanium screw in rabbit tibia

Abstract

Platelet-rich fibrin (PRF) is nowadays often used in various fields, but no removal torque studies have yet been done to evaluate the effects of the platelet-rich fibrin on the bone integration at the initial healing period. An experimental study have been performed in rabbits to evaluate whether the complete PRF clots can accelerate the bone integration of implants at the initial healing period after creating bone defects in tibias. The effec t of the complete PRF clots on bone integration was studied in two rabbit groups, 4-week group (group A) and 6-week group (group B) after preparing bony defects. Artificial bony defects were prepared in the tibias of rabbits. The complete PRF clots was applied to the defects in the experimental group, whereas the defects were unfilled in the control group. Four weeks later, machined implants were installed into the rabbit tibias (group A). Six weeks after installation, the removal torque from nine rabbits was measured to examine the bone healing effect of PRF. In another rabbits (group B), 6 weeks after preparing bony defects, installation was performed, and another 6 weeks after installation, the removal torque from nine rabbits was measured. The authors found a positive significant effect of the complete PRF clots on bone integration (higher bone density) in group A installed at 4 weeks after preparing bony defects (p?=?0.008; t-test), but not in group B installed at 6 weeks after preparing bony defects (p?=?0.677).