Platelet-to-neutrophil ratio predicts hemorrhagic transformation and unfavorable outcomes in acute ischemic stroke with intravenous thrombolysis

BACKGROUND Acute ischemic stroke(AIS)retains a notable stance in global disease burden,with thrombolysis via recombinant tissue plasminogen activator(rtPA)serving as a viable management approach,albeit with variable outcomes and the potential for complications like hemorrhagic transformation(HT).The platelet-to-neutrophil ratio(P/NR)has been considered for its potential prognostic value in AIS,yet its capacity to predict outcomes following rtPA administration demands further exploration.AIM To elucidate the prognostic utility of P/NR in predicting HT and clinical outcomes following intravenous rtPA administration in AIS patients.METHODS Data from 418 AIS patients treated with intravenous rtPA at Thammasat University Hospital from January 2018 to June 2021 were retrospectively analyzed.The relationship between P/NR and clinical outcomes[early neurological deterioration(E-ND),HT,delayed ND(D-ND),and 3-mo outcomes]was scrutinized.RESULTS Notable variables,such as age,diabetes,and stroke history,exhibited statistical disparities when comparing patients with and without E-ND,HT,D-ND,and 3-mo outcomes.P/NR prognostication revealed an optimal cutoff of 43.4 with a 60.3%sensitivity and a 52.5%specificity for 90-d outcomes.P/NR prognostic accuracy was statistically significant for 90-d outcomes[area under the curve(AUC)=0.562],D-ND(AUC=0.584),and HT(AUC=0.607).CONCLUSION P/NR demonstrated an association with adverse 3-mo clinical outcomes,HT,and D-ND in AIS patients post-rtPA administration,indicating its potential as a predictive tool for complications and prognoses.This infers that a diminished P/NR may serve as a novel prognostic indicator,assisting clinicians in identifying AIS patients at elevated risk for unfavorable outcomes following rtPA therapy.

Preparation and analysis of immunocompetence of recombinant fusion protein of the immunodominant region in chlamydial protease-like activity factor from Chlamydophila pneumoniae and its application in serodiagnosis

To clone the gene coding the immunodominant region in the chlamydial protease-like activity factor （CPAF） from Chlaroydophila pneumoniae, to analyze immunoeoropetenee of the expressed protein, and to evaluate its value in serodiagnosis, the CPAF immunodominant region gene was amplified, ligated into a pGEX6p-2 vector, and then the expressed recombinant protein was purified with glutathione Stransferase （GST） agarose gel FF after renaturation, then identified by SDS-PAGE and Western blot. A new indirect ELISA was developed with the purified protein as coating antigen. The immunogenicity of the recombinant protein was evaluated by immunization to New Zealand rabbits, and its immunoreactivity was analyzed by reacting with anti-C, pneumoniae antibody. 300 clinical sera samples were respectively detected by mieroimmunofluorescenee （MIF） as reference method and the indirect ELISA, and the difference between the two methods was analyzed. Cross-reactivity against Chlamydia trachomatis was investigated with the indirect ELISA to detect anti-C, trachomatis positive antisera. The results indicated that a 51.3 kDa recombinant protein was obtained. Western blot assay proved that the recombinant protein could merely specifically react with human anti- C. pneurnoniae antisera. The titers of the specific IgG antibodies in the immunized New Zealand rabbits were above 1 ： 16 000. Anti- C. pneumoniae IgG positive and negative reference sera were detected with the indirect ELISA, and the concordance rate of negative and positive results were both 100% （40/40）. The sensitivity and specificity of the indirect ELISA in comparison with MIF were 93.8% （45/48） and 100% （252/252） separately by detecting 300 clinical sera samples, and the concordance rate between the two methods was 99.0%. No cross reaction against C. trachomatis was found with the indirect ELISA to detect anti-C, trachomatis positive antisera. In conclusion, the prepared recombinant protein of the CPAF immunodominant region shows excellent immunocompetence and can be used to develop a new indirect ELISA as a method to detect anti-C, pneumoniae antibody for diagnosis of C. pneumoniae infection.

REGULATORY EFFECTS OF HUMAN RECOMBINANT IL-6 ON NATURAL KILLER CELL ACTIVITY OF HUMAN FETAL SPLEENS

In the present study it was proved first that human recombinant interleukin-6(HrIL-6) significantly augmented natural killer(NK) cell activity derived from human fetal spleens against K562 target cells in a 4 hours 51Cr release assay. The enhancement of NK activity with 24 hours preincubation in HrlL-6 was dose-dependent, and significantly higher than that of fresh NK cells and controls cultured with RPMI-1640 medium alone (P<0.001). We also found that IL-6 was able to augment NK activity from different fetal spleens at 20 to 40 weeks of gestation (up to 2.24 to 2.78 times), and no difference of NK activity of fetal splenocytes treated by HrIL-6 was observed between different fetal age (32.3% to 45.4%, P>0.05). Furthermore, IL-6-augmented NK activity of fetal splenocytes was very similar to adult levels (P>0.05). These finding strongly indicated that IL-6 plays an important role in the development of NK cell function during the gestational period, suggesting that IL-6 may be of importance in the regulation of host defense mechanisms against malignancies and viral diseases.

Expression of SNC73, a transcript of the immunoglobulin α-1 gene, in human epithelial carcinomas

AIM： To investigate the expression of SNC73, a transcript of the immunoglobulin α-1 gene （IgA1-H chain）, in human epitheliα-derived tumor cells. METHODS： Total RNAs and cell lysates were prepared from five different human epithelial cell lines derived from lung, stomach, liver, skin, and breast, respectively. RT-PCR and immunoblot analysis of these five cell lines were done. Both RT-PCR and immunochemistry were used to detect the expression of SNC73 in these cell lines. We also examined the expression of SNC73 in normal epithelial cells of colon mucosa by in situ hybridization. RT-PCR and immunoblot analysis were used to determine whether the recombination activating gene1/2 （RAG1 and RAG2） is present. The expression of three immunoglobulin transcription factors, EBF, E2A and Pax5, and the heavy chain of IgA1 and two types of light chains of immunoglobulin （κ and λ） in the aforementioned cell lines were analyzed by RT-PCR and immunochemistry, respectively. All the RT-PCR products were analyzed by sequencing. RESULTS： The results of RT-PCR and immunochemistry showed that both mRNA and protein of SNC73 were expressed in five human epitheliα-derived cancer cell lines. These data were further confirmed in the normal epithelial cells of colon mucosa by in situ hybridization. Also, the heavy chain of IgA1 and κ light chain were detected in these cells, but no λ light chain was observed. Both RAG1 and RAG2 were expressed in these human epitheliα-derived cancer cell lines and the sequence was identical to that expressed in pre-B and pre-T cells. In addition to RAG1 and RAG2, the mRNA in one of the immunoglobulin transcription factors, EBF, was also detected in these cell lines, and Pax5 was only expressed in SW480 cells, but no expression of E2A was observed in all the five cell lines. CONCLUSION： Immunoglobulin A1 is originally expressed and V（D）J recombination machine is also present in non-lymphoid cells, suggesting that V（D）J recombination machine mediates the assembly of immunoglobulin A1 in non-lymphoid cells as in prelymphocytes.

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.

Recombinant Tissue Plasminogen Activator-conjugated Nanoparticles Effectively Targets Thrombolysis in a Rat Model of Middle Cerebral Artery Occlusion

The efficacy and safety of recombinant tissue plasminogen activator （rtPA） need to be improved due to its low bioavailability and requirement of large dose administration. The purpose of this study was to develop a fibrin-targeted nanoparticle （NP） drug delivery system for thrombosis combination therapy. We conjugated rtPA to poly（ethylene glycol）- poly（ε-caprolactone） （PEG-PCL） nanoparticles （rtPA-NP） and investigated its physicochemical characteristics such as particle size, zeta potential, enzyme activity of conjugated rtPA and its storage stability at 4℃. The thrombolytic activity of rtPA-NP was evaluated in vitro and in vivo as well as the half-life of rtPA-NP, the properties to fibrin targeting and its influences on systemic hemostasis in vivo. The results showed that rtPA-NP equivalent to 10% of a typical dose of rtPA could dissolve fibrin clots and were demonstrated to have a neuroprotective effect after focal cerebral ischemia as evidenced by decreased infarct volume and improved neurological deficit （P〈0.001）. RtPA-NP did not influence the in vivo hemostasis or coagulation system. The half-life of conjugated rtPA was shown to be approximately 18 times longer than that of free rtPA. These experiments suggested that rtPA-conjugated PEG-PCL nanoparticles might be a promising fibrin-targeted delivery system for a combination treatment of thrombosis.

Therapeutic effect of recombinant tissue plasminogen activator on acute cerebral infarction at different times

BACKGROUND:The study aimed to compare the therapeutic effect of recombinant tissue plasminogen activator(rt-PA) on the onset of acute cerebral infarction(ACI) at different time points of the first 6 hours.METHODS:A retrospective analysis was conducted in 74 patients who received rt-PA thrombolysis treatment within 4.5 hours after ACI and another 15 patients who received rt-PA thrombolysis treatment between 4.5-6 hours after ACI.RESULTS:National Institute of Health Stroke Scale(NIHSS) scores were statistically decreased in both groups(P>0.05) at 24 hours and 7 days after ACI.There was no significant difference in modified ranking scores and mortality at 90 days after the treatment between the two groups(P>0.05).CONCLUSIONS:The therapeutic effect and mortality of rt-PA treatment in patients with ACI between 4.5-6 hours after the onset of the disease were similar to those in patients who received rtPA within 4.5 hours after the onset of this disease.Therefore,intravenous thrombolytic therapy for ACI within 4.5-6 hours after ACI was effective and safe.

Minimally invasive puncture and drainage or patients with hypertensive spontaneous basal ganglia intracerebral hemorrhage: A prospective non-randomized comparative study of 198 cases

Background: The treatment of hypertensive spontaneous intracranial hemorrhage(ICH) is still controversial. The purpose of the present study was to investigate whether minimally invasive puncture and drainage(MIPD) could provide improved patient outcome compared with decompressive craniectomy(DC).Methods: Eligible, consecutive patients with ICH(≥30 ml, in basal ganglia, within 24 hours of ictus) were nonrandomly assigned to receive MIPD(group A) or to undergo DC(group B) hematoma evacuation. The primary outcome was death at 30 days after onset. Functional independence was assessed at 1 year using the Glasgow Outcome Scale(GOS, scores range from 1 to 5, score 1 indicating death, ≥4 indicating functional independence, with lower scores indicating greater disability). Results: A total of 198 patients met the per protocol analysis(84 cases in group A and 114 cases in group B), including 9 cases lost during follow-up(2 cases in group A and 7 cases in group B). For these 9 patients, their last observed data were used as their final results for intention-to-treat analysis. The mean age of all patients was 57.1 years(range of 31-95 years), and 114 patients were male. The initial Glasgow Coma Scale(GCS) score was 8.1±3.4, and the National Institutes of Health Stroke Scale(NIHSS) score was 20.8±5.3. The mean hematoma volume(HV) was 56.7±23.0 ml(range of 30-144 ml), and there was extended intraventricular hemorrhage(IVH) in 134 patients(67.7%). There were no significant intergroup differences in the above baseline data, except group A had a higher mean age(59.4±14.5years) than the mean age of group B(55.3±11.1 years, P=0.025). The total cumulative mortalities at 30 days and 1 year were 32.3% and 43.4%, respectively, and there were no significant differences between groups A and B(30 days: 27.4% vs. 36.0%, P=0.203; 1 year: 36.1% vs. 48.2%, P=0.112, respectively). However, the mortality for patients ≤60 years, NIHSS【15 or HV≤60 ml was significantly lower in group A than that in group B(all P【0.05). The total cumulative functional independence at 1 year was 26.8%, and the difference between group A(33/43, 39.3%) and group B(20/144, 17.5%) was significant(absolute difference 21.7%, odds ratio [OR] 0.329, 95% confidence interval [CI] 0.171 to 0.631, P=0.001). For patient with severe IVH, the 30 days and 1 year mortality rates were significant lower in group B than those in group A(P=0.025, P=0.036). However, the number of favorable outcomes had no significant difference between groups at 1 year post ictus. Multivariate logistic regression analysis showed that a favorable outcome after 1 year was associated with the difference in therapies(OR 0.280, 95% CI 0.104–0.752, P=0.012), age(OR 0.215, 95% CI 0.069–0.671, P=0.008), GCS(OR 1.187, 95% CI 1.010–1.395, P=0.037), HV(OR 0.943, 95% CI 0.906–0.982, P=0.005), IVH(OR 0.655, 95% CI 0.506–0.849, P=0.001) and PI(OR 0.211, 95% CI 0.071–0.624, P=0.001). Conclusions: Our results suggest that for patients with hypertensive spontaneous ICH(HV≥30 ml in basal ganglia), MIPD may be a more effective treatment than DC, as assessed by a higher rate of functional independence at 1 year after onset as well as reduced mortality in patients ≤60 years of age, NIHSS【15 or HV≤60 ml. For patients with HV 】60 ml, deep coma and severe IVH, the outcomes of the two therapies were similar.

Advances in surgical treatment of intraventricular hemorrhage

Intraventricular hemorrhage(IVH)is a dangerous acute cerebrovascular disease.The 30-day mortality rate of patients with IVH is 5 times that of patients with cerebral hemorrhage.In recent years,more and more studies have shown that timely clearance of intraventricular hematoma is closely related to good prognosis.The advantages of surgical intervention in patient management are impressive.This article briefly reviews the positive advances in the surgical treatment of IVH such as external ventricular drainage(EVD)combined with intraventricular fibrinolysis(IVF)and continuous lumbar cistern drainage(LCD),as well as their safety and efficacy,especially the remarkable role of neuroendoscopy in the removal of intraventricular hematomas.

Submacular hemorrhage:treatment update and remaining challenges

Submacular haemorrhage(SMH)is a sight threatening complication that can occur in exudative age related macular degeneration(AMD),but has been described to occur more frequently in eyes with polypoidal choroidal vasculopathy(PCV).Left untreated,SMH carries a grave visual prognosis.Thus,expedient diagnosis and effective management of this complication is of paramount importance.The treatment strategies for SMH include(I)displacement of blood from the fovea,usually by injection of an expansile gas;(II)pharmacologic clot lysis such as with recombinant tissue plasminogen activator(rtPA);and(III)treatment of the underlying choroidal neovascularization(CNV)or PCV,such as with anti-vascular endothelial growth factor(anti-VEGF)agents.These three strategies have been employed in isolation or in combination,some concurrently and others in stages.rtPA has demonstrable effect on the liquefaction of submacular clots but there are remaining uncertainties with regards to the dose,safety and the timing of initial and repeat treatments.Potential side effects of rtPA include retinal pigment epithelial toxicity,increased risk of breakthrough vitreous haemorrhage and systemic toxicity.In cases presenting early,pneumatic displacement alone with anti-VEGF may be sufficient.Anti-VEGF monotherapy is a viable treatment option particularly in patients with thinner SMH and those who are unable to posture post pneumatic displacement.

Factors influencing clinical outcomes of acute ischemic stroke treated with intravenous recombinant tissue plasminogen activator

Background Thrombolysis with recombinant tissue plasminogen activator （rt-PA） has gained international recognition, clinical outcomes following this thrombolytic therapy varied from patient to patient. Factors affecting clinical outcomes have not been well understood yet, so this retrospective case-control study aimed to investigate factors that may influence clinical outcomes of acute ischemic stroke treated with intravenous rt-PA. Methods One hundred and one patients with acute ischemic stroke who received intravenous rt-PA thrombolysis within 4.5 hours from disease onset were included. Patients were divided into good or poor outcome group according to modified Rankin Scale （mRS） score, good outcome group： mRS score of 0-1; poor outcome group： mRS of 2-6. Stroke characteristics were compared between the two groups. Factors for stroke outcomes were analyzed via univariate analysis and Logistic regression. Results Of the 101 patients studied, patients in good outcome group （n=55） were significantly younger than patients in poor outcome group （n=46, （62.82±14.25） vs. （68.81±9.85） years, P=0.029）. Good outcome group had fewer patients with diabetic history （9.09% vs. 28.26%, P=0.012）, fewer patients with leukoaraiosis （7.27% vs. 28.26%, P=-0.005） and presented with lower blood glucose level （（5.72±1.76） vs. （6.72±1.32） mmol/L, P=0.012）, lower systolic blood pressure level （（135.45±19.36） vs. （148.78±19.39） mmHg, P=0.003）, lower baseline NIHSS score （12.02±5.26 vs. 15.78±4.98, P=0.002） and shorter onset-to-treatment time （OTT） （（2.38±1.21） vs. （2.57±1.03） hours, P=0.044） than poor outcome group. Logistic regression analysis showed that absence of diabetic history （odds ratio （OR） 0.968 （95% CI 0.941-0.996））, absence of leukoaraiosis （OR 0.835 （95% C/0.712-0.980））, lower baseline NIHSS score （OR 0.885 （95% Cl 0.793- 0.989））, lower pre-thrombolysis systolic blood pressure （OR 0.962 （95% CI 0.929-0.997））, and lower blood glucose level （OR 0.699 （95% Cl 0.491-0.994）） before thrombolysis were significantly associated with better outcome. Conclusion Patients with no history of diabetes, no leukoaraiosis, low blood glucose level, low systolic blood pressure level and low baseline NIHSS score before thrombolvsis have a better outcome.

Analysis of the Efficacy and Safety of Recombinant Tissue Plasminogen Activator for Chinese Patients over 80 Years of Age with Acute Ischemic Stroke： A Pilot Study

Dear Editor,Recombinant tissue plasminogen activator（rt-PA）remains the only proven therapy for acute ischemic stroke patients who meet the selection criteria for treatment[1–3].A small number of studies have investigated the efficacy of rt-PA for elderly stroke patients in Japanese and Caucasian populations[4,5].According to the American Heart Association/American Stroke Association Guidelines,

Factors Associated with Thrombolysis Outcome in Ischemic Stroke Patients with Atrial Fibrillation

The outcome of early intravenous thrombolysis for ischemic stroke in patients with atrial fibrillation（AF）is worse than that without thrombosis. How to increase the efficacy of intravenous thrombolysis for AF-related ischemic stroke remains largely unknown. In this study, we investigated factors that influence the effect of intravenous thrombolysis in these patients. Our results showed that thrombolysis was independently associated with a favorable outcome（P ／ 0.001） and did not influence the mortality of AF-related ischemic stroke, although it increased the risk of hemorrhage within 24 h after treatment. Risk factors for a poor outcome at admission were：heart failure（P = 0.045）; high systolic pressure（P = 0.039）; high blood glucose（P = 0.030）; and a high National Institutes of Health Stroke Scale（NIHSS） score（P ／ 0.001）. Moreover, high systolic pressure at admission（P = 0.007）, high blood glucose（P = 0.027）, and a high NIHSS score（P ／ 0.001） were independent risk factors for mortality at 3 months. Besides thrombolysis, a high NIHSS score（P = 0.006） and warfarin taken within 48 h before stroke onset（P = 0.032） were also independent risk factors for symptomatic hemorrhage within 24 h after treatment. Ischemic stroke patients with AF benefited from intravenous thrombolysis with recombinant tissue plasminogen activator within 4.5 h after stroke.

An Experimental Study on the Flexibility of Prevention against Thrombosis Following Mechanical Valve Replacement by tPA Gene Transduction

Objectives Use a gene suture immersed recombinant tissue-type plasminogen activator （r-tPA）expression plasmid to transduce myocardia to prevent the thrombosis after mechanical tricuspid valve replacement in pigs. Methods A r-tPA gene plasmid was constructed and conjugated to a novel cationic phosphonolipid and a r-tPA gene suture was made. Eighteen pigs were selected and divided into two groups at randomization. There were 9 pigs in the experimental group and 9 in the control group, all the 18 pigs＇ tricuspids were replaced with mechanical valves. The gene threads were sutured into the right ventricular walls near mechanical valves and an ultrasound was used on the surfaces of the right ventricular walls for the gene transfer in the experimental group. Coagulative function, D-dimer level of the blood and the thrombosis on the surfaces of the valves were observed. Results r-tPA gene plasmid was successfully constructed and r-tPA protein was expressed in the ventricular cells around the gene sutures. D-dimer reached its peak level （ 1.67 ±0. 79） μg · mL^-1 in 1 week after operation in two groups, but it decreased to preoperation level thereafter in control group and kept on the high level and reincreased to a new high level （ 1.89 ± 0.79 ） μg · mL^-1 until the end of the experiment in experimental group. The thromboses around the valves were found in all the control group （100%） but only 1 （ 11.11% ） case in experimental group. There were no changes in prothrombin time pre and post operation in two groups. Conclusions Using gene suture immersed r-tPA expression plasmid to transduce myocardia might be a best substitution for life long anti-coagulation therapy for the patients, who underwent operation.