CRM 197(cross-reacting material 197),a naturally occurring mutant of diphtheria toxin,is a safe and effective vaccine vector and extensively used on developing conjugate or combined vaccines.The mutant loses its enzymatic activity,but fully retains its receptor-binding ability and immunogenicity.In current work,the diphtheria toxin mutant CRM 197 and its fusion proteins with the receptor-binding do-main of botulinum neurotoxin serotype E(EHc)were developed using genetic engineering technology.These recombinant proteins were confirmed by Western blotting and SDS-PAGE.BALB/c mice were im-munized with the CRM197-EHc and EHc-CRM197 fusion proteins,and their immunogenicity was evalua-ted.These two fusion protein molecules,CRM197-EHc and EHc-CRM197,as subunit vaccines,elicited a robust humoral immune response targeting both CRM197 and EHc antigens in the immunized mice.Compared to the mixture of CRM197 and EHc,the mice vaccinated with the fusion proteins(CRM197-EHc and EHc-CRM197)induced higher levels of anti-CRM197 antibodies,and the mice vaccinated with EHc-CRM197 also generated strongest anti-EHc antibodies.Consequently,as a carrier molecule in the fusion protein vaccine,EHc enhances the immunogenicity of CRM197 molecules.Likewise,CRM197 boosts the immunogenicity of EHc in the EHc-CRM197 fusion protein.

Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer(BC)and esophageal cancer(EC)tissue samples in order to establish evidence-based protocols for biobank sample management.Methods The tumor(T)and paired normal(N)tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank,Shantou Central Hospital.Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis.The samples were divided into two groups of<15 min and 15-30 min according to ischemia time,and RNA quality was analyzed at 4 storage periods of 8-10 months(T1),14-16 months(T2),26-28 months(T3)and 38-40 months(T4).Results In 96 analyzed samples,93.8%(90/96)exhibited high quality(RIN≥6),with 89.6%(43/48)in BC and 97.9%(47/48)in EC.Significant differences in RIN were observed between BC group and EC group(8.050 vs.8.600,P=0.009).In EC group,RIN value was significantly negatively correlated with RNA yield(P<0.001).Moreover,RIN values of tumor-normal pairs exhibited markedly significant differences(7.550 vs.9.000,P<0.001).In contrast,no significant difference was detected in BC group(8.200 vs.7.700,P=0.348).Statistical analysis showed that RIN value was positively correlated with 28S/18S(P<0.001),but had no correlation with tumor content(P=0.676)and necrotic content(P=0.055).Neither ischemia time(<15 min vs.15-30 min:8.200 vs.8.300,P=0.932)nor storage periods(T1-T4:8.400,7.700,8.450,8.600,P=0.163)compromised RNA quality.Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples.However,limited ischemia time(≤30 min)and long-term storage period(38-40 months)do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

Sampling is the first stage of the DNA Typing in forensic science,and some studies found that the sampling technology affected the outcomes of"Contact DNA"testing.Since swabs are the most commonly used collection instrument for"Contact DNA",the researchers in forensic science field should pay more attention to study the collection efficiency of swabs.However,currently the majority of relevant research is focused on downstream testing processes,such as typing and amplification;the corresponding work on sampling performance by swabs is quite limited.Furthermore,the absence of an internationally recognized standard procedure makes it impossible to directly compare the collection results among laboratories.In this paper,a review on the use of swabs in the forensic science field is presented.It analyzes the collection efficiency,advantages and disadvantages of different swabs,and investigates the need and feasibility of developing a standardized method to evaluate the swab collection efficiency,for improving the sampling performance of swabs and ultimately increasing the detection rate of"Contact DNA"in forensic science.

CRM 197(cross-reacting material 197),a naturally occurring mutant of diphtheria toxin,is a safe and effective vaccine vector and extensively used on developing conjugate or combined vaccines.The mutant loses its enzymatic activity,but fully retains its receptor-binding ability and immunogenicity.In current work,the diphtheria toxin mutant CRM 197 and its fusion proteins with the receptor-binding do-main of botulinum neurotoxin serotype E(EHc)were developed using genetic engineering technology.These recombinant proteins were confirmed by Western blotting and SDS-PAGE.BALB/c mice were im-munized with the CRM197-EHc and EHc-CRM197 fusion proteins,and their immunogenicity was evalua-ted.These two fusion protein molecules,CRM197-EHc and EHc-CRM197,as subunit vaccines,elicited a robust humoral immune response targeting both CRM197 and EHc antigens in the immunized mice.Compared to the mixture of CRM197 and EHc,the mice vaccinated with the fusion proteins(CRM197-EHc and EHc-CRM197)induced higher levels of anti-CRM197 antibodies,and the mice vaccinated with EHc-CRM197 also generated strongest anti-EHc antibodies.Consequently,as a carrier molecule in the fusion protein vaccine,EHc enhances the immunogenicity of CRM197 molecules.Likewise,CRM197 boosts the immunogenicity of EHc in the EHc-CRM197 fusion protein.

Objective:To explore the differences in the whole-brain functional connectivity characteristics of magnetoencephalography gamma sub-band in resting state between patients with depression with high and low suicidal ideation.Methods:Clinical data of 60 patients with depression (depression group) who visited the Department of Psychiatry, Nanjing Brain Hospital Affiliated to Nanjing Medical University from October 2021 to June 2024 were prospectively collected, including 20 males and 40 females aged 16-41 years. During the same period, 75 healthy controls (healthy control group) were recruited, matched for gender and age, comprising 33 males and 42 females, aged 18-43 years. The Hamilton Depression Scale-17 items (HAMD-17) was used to assess the degree of depression in patients, and the score of the third item was used to divide the patients into a high suicidal ideation group of 37 cases (2-3 points) and a low suicidal ideation group of 23 cases (0-1 points). The Beck Scale for Suicide Ideation-Chinese Version (BSI-CV) was used to assess the intensity of suicidal ideation in patients in the last week and at the most severe stage. Brain imaging data of the subjects were collected using a CTF-275 channel magnetoencephalography device and a 3 T magnetic resonance imaging device. The functional connectivity matrix of the gamma sub-band depression group, healthy control group, high suicidal ideation group, and low suicidal ideation group was calculated using the envelope correlation method. The network-based statistics (NBS) algorithm was used to correct for multiple comparisons and compare the differences in brain network functional connectivity between the depression group/healthy control group and between the high/low suicidal ideation groups.Results:Compared with the healthy control group, the functional connectivity between the bilateral middle frontal gyrus, right inferior frontal gyrus orbital part, right anterior cingulate gyrus, left middle frontal gyrus orbital part, and right superior parietal lobule in the depression group was significantly enhanced in the low gamma frequency band (30-60 Hz) ( θ=3, P=0.042). Compared with the low suicidal ideation group, the functional connectivity between the right amygdala as a central node and the right superior frontal gyrus, left hippocampus, right upper temporal pole, and left transverse temporal gyrus in the high suicidal ideation group was significantly enhanced in the high gamma frequency band (60-90 Hz) ( θ=3, P=0.049). Conclusion:In the low gamma frequency band, patients with depression have neural dysfunction mainly in the frontal lobe. In the high gamma frequency band, patients with high suicidal ideation have functional connectivity abnormalities centered on the amygdala.

Objective:To evaluate the efficacy and safety of intrasaccular flow disruptor in wide-necked intracranial aneurysms.Methods:One hundred and seventeen patients with wide-necked intracranial aneurysms treated with intrasaccular flow disruptor were collected from Department of Neurointervention (First Affiliated Hospital of Zhengzhou University), Department of Neurosurgery (Beijing Tiantan Hospital, Capital Medical University), Department of Cerebrovascular Surgery, Neurosurgery Center (Zhujiang Hospital, Southern Medical University), and Department of Neurosurgery (First Affiliated Hospital of Harbin Medical University) from August 2022 to March 2024. Raymond-Roy Occlusion Classification (RROC) was employed to evaluate aneurysm embolization immediately after procedure; cranial CT or MRI within 48 hours of embolization were performed to identify any new intracranial hemorrhage, subarachnoid hemorrhage, or new symptomatic cerebral infarction related to the intracranial aneurysms. Modified Rankin Scale (mRS) was used to assess the neurological function at discharge. Imaging follow-up and outpatient follow-up were performed at 6 months after embolization to evaluate the aneurysm occlusion degree and complications.Results:A total of 117 intrasaccular flow disruptors were implanted in 117 patients, with a technical success rate of 100%; 115 patients (98.3%) enjoyed successful one-time release of their disruptors, and 2 patients (1.7%) required retrieval and redirection of the disruptors before second successful attempt. Flow disruptor plus stent was performed in 13 patients (11.1%). Immediately after procedure, RROC grading I was noted in 3 patients, grading II in 51 patients and grading III in 63 patients. Cranial CT or MRI within 48 hours of embolization indicated no new intracranial hemorrhage, subarachnoid hemorrhage, or symptomatic cerebral infarction related to the intracranial aneurysms. All patients had mRS score of 0 at discharge. Eighty-three patients completed a 6-month follow-up (RROC grading I in 41 patients, grading II in 33 patients and grading III in 9 patients), without ischemic or hemorrhagic adverse events.Conclusion:The results of this study preliminarily suggest that intrasaccular flow disruptor is effective and safe in wide-necked intracranial aneurysms.

Objective:To evaluate the efficacy and safety of Neuroform Atlas stent-assisted coil embolization in patients with middle cerebral artery bifurcation aneurysms.Methods:A retrospective analysis was performed; the clinical data of 46 patients with middle cerebral artery bifurcation aneurysms accepted Neuroform Atlas stent-assisted coil embolization in First Affiliated Hospital of Zhengzhou University, Beijing Tiantan Hospital Affiliated to Capital Medical University, First Affiliated Hospital of Harbin Medical University, Zhujiang Hospital of Southern Medical University and First Affiliated Hospital of Chongqing Medical University from January 2022 to March 2024 were collected. There were 28 ruptured aneurysms (60.87%) and 18 unruptured aneurysms (39.13%). Follow-up was performed for more than 3 months; Raymond-Roy grading was used to evaluate the aneurysm embolization immediately after embolization and during follow-up; perioperative hemorrhagic or ischemic complications were recorded; modified Rankin Scale (mRS) was used to evaluate the prognosis of the patients at discharge and during follow-up (mRS score≤2: good prognosis, and mRS score>2: poor prognosis).Results:Coil embolization was successful in all 46 patients. DSA immediately after embolization showed that 41 patients (89.13%) had completely occluded aneurysms (Raymond-Roy grading I), 2 patients (4.35%) had residual aneurysm neck (Raymond-Roy grading Ⅱ) and 3 patients (6.52%) had partially occluded aneurysms (Raymond-Roy grading Ⅲ). Perioperative complications occurred in 5 patients, including 2 with postoperative cerebral infarction, 1 with hydrocephalus, 1 with postoperative pneumonia leading to respiratory failure, and 1 with stent thrombosis during embolization. Both at discharge and 3 months after embolization, 43 patients (93.48%) had good prognosis and 3 patients (6.52%) had poor prognosis. No obvious ischemic complications (such as stent restenosis) or hemorrhagic complications (such as re-rupture of the aneurysms) were found in all patients. Thirty patients (65.22%) had imaging follow-up for 6-12 months: 26 (86.67%) had Raymond-Roy grading I, 3 (10.00%) had Raymond-Roy grading II, and 1 (3.33%) had Raymond-Roy grading III.Conclusion:Neuroform Atlas stent-assisted coil embolization has good short-term efficacy and high safety in middle cerebral artery bifurcation aneurysms, but long-term follow-up observation is still needed to verify its efficacy.

Sampling is the first stage of the DNA Typing in forensic science,and some studies found that the sampling technology affected the outcomes of"Contact DNA"testing.Since swabs are the most commonly used collection instrument for"Contact DNA",the researchers in forensic science field should pay more attention to study the collection efficiency of swabs.However,currently the majority of relevant research is focused on downstream testing processes,such as typing and amplification;the corresponding work on sampling performance by swabs is quite limited.Furthermore,the absence of an internationally recognized standard procedure makes it impossible to directly compare the collection results among laboratories.In this paper,a review on the use of swabs in the forensic science field is presented.It analyzes the collection efficiency,advantages and disadvantages of different swabs,and investigates the need and feasibility of developing a standardized method to evaluate the swab collection efficiency,for improving the sampling performance of swabs and ultimately increasing the detection rate of"Contact DNA"in forensic science.

Objective To compare the predictive effectiveness of the Glasgow coma scale(GCS),GCS-pupils scale(GCS-P),Glasgow-Pittsburgh coma scale(GPCS),full outline of unresponsiveness scale(FOUR),and coma recovery scale-revised(CRS-R)in forecasting the prognosis of severe stroke patients.Methods A prospective,consecutive cohort of severe stroke patients admitted to the Department of Neurology,First Medical Center of Chinese PLA General Hospital from September 2021 to April 2024 was enrolled.Demographic and clinical data were collected,including age,sex,length of hospital stay,diagnosis(severe ischemic stroke,severe cerebral hemorrhage,aneurysmal subarachnoid hemorrhage),medical history(hypertension,diabetes,coronary artery disease),smoking and drinking habits,vital signs upon admission(temperature,pulse,respiration,blood pressure),neurological examination findings(including speech and brainstem reflexes)at admission,head imaging results(CT,MRI)within 24 h of admission to assess the presence of brain herniation,and whether intubation occurred within 24 h of admission.Patients underwent GCS,GCS-P,GPCS,FOUR,and CRS-R scoring within 8h of admission.Telephone follow-up was conducted at 6 months post-stroke to assess outcomes using the modified Rankin scale(mRS),with mRS scores of 0-2 classified as the good prognosis group and 3-6 as the poor prognosis group.The receiver operating characteristic(ROC)curve was used to assess the prognostic prediction value of the five scales for poor outcomes at 6 months.The area under the ROC curve(AUC)was calculated,and pairwise comparisons of AUC were performed using the Delong test.Results A total of 179 severe stroke patients were enrolled,including 116 males and 63 females.The group consisted of 132 patients with severe ischemic stroke,30 with severe intracerebral hemorrhage,and 17 with aneurysmal subarachnoid hemorrhage.At 6months,126patients had a poor prognosis and 53 had a good prognosis.(1)There were statistically significant differences in age,temperature,pulse,history of coronary artery disease,smoking and drinking habits,presence of speech impairment,abnormal brainstem reflexes,brain herniation,intubation within 24 h of admission,and GCS,GCS-P,GPCS,FOUR,and CRS-R scores between the poor and good prognosis groups(all P＜0.05).(2)ROC analysis revealed that the AUC(95％CI)for predicting poor outcomes at 6 months in severe stroke patients for GCS,GCS-P,GPCS,FOUR,and CRS-R were 0.808(0.742-0.863),0.815(0.750-0.869),0.828(0.765-0.880),0.841(0.780-0.892),and 0.831(0.768-0.883),respectively.Sensitivities were 76.98％,78.57％,82.54％,84.13％,and 82.54％,and specificities were 73.58％,73.58％,67.92％,71.70％,and 73.58％,respectively.The FOUR had the highest AUC,with an optimal cutoff value of 13.(3)Pairwise comparisons of AUC showed a statistically significant difference between the FOUR and GCS(the difference value of AUC is 0.034,95％CI 0.004-0.064,Z=2.194,P=0.028),but no significant differences were observed between other scales(all P＞0.05).Conclusion Compared to GCS,GCS-P,GPCS,and CRS-R,FOUR may provide more valuable prognostic information for severe stroke patients.

Ureaplasma urealyticum (UU) is one of the most commonly occurring pathogens associated with genital tract infections in infertile males, but the impact of seminal UU infection in semen on intrauterine insemination (IUI) outcomes is poorly understood. We collected data from 245 infertile couples who underwent IUI at The First Affiliated Hospital of USTC (Hefei, China) between January 2021 and January 2023. The subjects were classified into two groups according to their UU infection status: the UU-positive group and the UU-negative group. We compared semen parameters, pregnancy outcomes, and neonatal birth outcomes to investigate the impact of UU infection on IUI outcomes. There were no significantly statistical differences in various semen parameters, including semen volume, sperm concentration, total and progressive motility, sperm morphology, leukocyte count, the presence of anti-sperm antibody, and sperm DNA fragmentation index (DFI), between the UU-positive and UU-negative groups of male infertile patients (all P > 0.05). However, the high DNA stainability (HDS) status of sperm differed between the UU-positive and UU-negative groups, suggesting that seminal UU infection may affect sperm nuclear maturation ( P = 0.04). Additionally, there were no significant differences in pregnancy or neonatal birth outcomes between the two groups (all P > 0.05). These results suggest that IUI remains a viable and cost-effective option for infertile couples with UU infection who are facing infertility issues.
Humans ; Male ; Ureaplasma Infections/complications* ; Female ; Infertility, Male/therapy* ; Ureaplasma urealyticum/isolation & purification* ; Pregnancy ; Adult ; Pregnancy Outcome ; Semen Analysis ; Insemination, Artificial ; Semen/microbiology* ; China