Objective To investigate the impact of different cell digestion methods on the binding of a humanized monoclonal antibody targeting CD3L1(designated as 5H)to tumor cells.Methods Trypsin solution and a collagenase/neutral protease cocktail,combined with two digestion temperatures(room temperature and 37℃),were used to dissociate,passage,and collect various adherent tumor cell lines.Flow cytometry and confocal microscopy were then employed to compare differences in 5H antibody binding under different cell digestion conditions.Results Cells dissociated and passaged with trypsin solution at room temperature exhibited only weak binding to the 5H antibody,whereas those treated with the collagenase/neutral protease cocktail at room temperature showed significantly stronger binding.There were notable differences in the binding signal and the proportion of antibody-stained positive cells between the two groups(P<0.01).Additionally,cells treated with the collagenase/neutral protease cocktail exhibited antibody binding signals that were dependent on the concentration of the 5H antibody,enabling receptor occupancy analysis.Compared to digestion at room temperature,treatment at 37℃ resulted in a slight reduction in the binding of target cells to the 5H antibody,however,this difference was not statistically significant.Conclusion Trypsin digestion significantly reduces the binding of the anti-CD3L1 antibody to tumor cells,whereas the collagenase/neutral protease cocktail effectively preserves it.With an appropriate choice of digestion enzymes,the impact of digestion temperature is relatively minor.These findings suggest that optimized cell digestion protocols should be employed when studying interactions between antibody drugs and cell surface antigens.

Objective To investigate the impact of different cell digestion methods on the binding of a humanized monoclonal antibody targeting CD3L1(designated as 5H)to tumor cells.Methods Trypsin solution and a collagenase/neutral protease cocktail,combined with two digestion temperatures(room temperature and 37℃),were used to dissociate,passage,and collect various adherent tumor cell lines.Flow cytometry and confocal microscopy were then employed to compare differences in 5H antibody binding under different cell digestion conditions.Results Cells dissociated and passaged with trypsin solution at room temperature exhibited only weak binding to the 5H antibody,whereas those treated with the collagenase/neutral protease cocktail at room temperature showed significantly stronger binding.There were notable differences in the binding signal and the proportion of antibody-stained positive cells between the two groups(P<0.01).Additionally,cells treated with the collagenase/neutral protease cocktail exhibited antibody binding signals that were dependent on the concentration of the 5H antibody,enabling receptor occupancy analysis.Compared to digestion at room temperature,treatment at 37℃ resulted in a slight reduction in the binding of target cells to the 5H antibody,however,this difference was not statistically significant.Conclusion Trypsin digestion significantly reduces the binding of the anti-CD3L1 antibody to tumor cells,whereas the collagenase/neutral protease cocktail effectively preserves it.With an appropriate choice of digestion enzymes,the impact of digestion temperature is relatively minor.These findings suggest that optimized cell digestion protocols should be employed when studying interactions between antibody drugs and cell surface antigens.

Objective: To observe the incidence and treatment of radiation rectal injury complicated with anxiety, depression and somatic symptom disorder. Methods: A cross-sectional survey research method was carried out. Patients with radiation rectal injury managed by members of the editorial board of Chinese Journal of Gastrointestinal Surgery were the subjects of investigation. The inclusion criteria of the survey subjects: (1) patients suffered from pelvic tumors and received pelvic radiotherapy; (2) colonoscopy showed inflammatory reaction or ulcer in the rectum. Exclusion criteria: (1) patient had a history of psycho-somatic disease before radiotherapy; (2) patient was unable to use a smart phone, unable to read and understand the questions in the questionnaire displayed on the phone; (3) patient refused to sign an informed consent form. According to the SOMA self-rating scale, PHQ-15 self-rating scale, GAD-7 and PHQ-9 self-rating scale, the electronic questionnaire of "Psychological Survey of Radiation Proctitis" was designed. The questionnaire was sent to patients with radiation rectal injury managed by the committee through the WeChat group. Observational indicators: (1) radiation rectal injury symptom assessment: using SOMA self-rating scale, radiation rectal injury symptom classification: mild group (≤3 points), moderate group (4-6 points) and severe group (> 6 points); (2) incidence of anxiety, depression and physical disorder: using GAD-7, PHQ-9 and PHQ-15 self-rating scales respectively for assessment; (3) correlation of radiation rectal injury symptom grading with anxiety, depression, and somatic symptom disorder. Results: Seventy-one qualified questionnaires were collected, of which 41 (56.9%) were from Guangzhou. Among the 71 patients, 6 were males and 65 were females; the mean age was (55.7±9.3) years old and 48 patients (67.6%) were less than 60 years old; the median confirmed duration of radiation rectal injury was 2.0 (1.0, 5.0) years. (1) Evaluation of symptoms of radiation rectal injury: 18 cases of mild (25.4%), 27 cases of moderate (38.0%), and 26 cases of severe (36.6%). (2) Incidence of anxiety, depression and somatic disorder: 12 patients (16.9%) without comorbidities; 59 patients (83.1%) with anxiety, depression, or somatic disorder, of whom 2 patients only had anxiety, 1 patient only had depression, 9 only had somatic disorder, 2 had anxiety plus depression, 4 had anxiety plus somatic disorder, 2 had depression plus somatic disorder, and 40 had all three symptoms. (3) correlation of radiation rectal injury grading with anxiety, depression, and somatic symptom disorder: as compared to patients in mild group and moderate group, those in severe group had higher severity of anxiety and somatic symptom disorder (Z=-2.143, P=0.032; Z=-2.045, P=0.041), while there was no statistically significant difference of depression between mild group and moderate group (Z=-1.176, P=0.240). Pearson correlation analysis revealed that radiation rectal injury symptom score was positively correlated with anxiety (r=0.300, P=0.013), depression (r=0.287, P=0.015) and somatic symptom disorder (r=0.344, P=0.003). Conclusions: The incidence of anxiety, depression, and somatic symptom disorder in patients with radiation rectal injury is extremely high. It is necessary to strengthen the diagnosis and treatment of somatic symptom disorder, so as to alleviate the symptoms of patients with pelvic perineum pain and improve the quality of life.
Aged ; Anxiety ; Cross-Sectional Studies ; Depression ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Rectum ; Surveys and Questionnaires

Objective To investigate the prevalence and influencing factors of Enterobius vermicularis infections among children in Fanxian County, Henan Province in 2019, so as to provide insights into the management of enterobiasis. Methods Five kindergartens were selected in urban and rural areas of Fanxian County, Henan Province using the stratified sampling method in 2019, and a census of E. vermicularis infections was performed among all children in the kindergartens. E. vermicularis eggs were detected using adhesive and scotch cellophane-tape anal swab methods, and the basic characteristics of children and their families, health habits and the kindergartens’ information were investigated with questionnaires. Logistic regression analysis was used to investigate the risk factors and protective factors of pinworm infection in children. Results A total of 671 children were tested, and the mean prevalence of E. vermicularis infections was 15.50% (104/671). The prevalence of E. vermicularis infections was higher among children in rural kindergartens (28.13%, 72/256) than in urban kindergartens (7.71%, 32/415) (χ² = 50.380, P < 0.01), and greater in private kindergartens (32.26%, 60/186) than in public kindergartens (9.07%, 44/485) (χ² = 55.183, P < 0.01). There was no gender-specific prevalence of E. vermicularis infections among children (χ² = 1.442, P > 0.05), and the prevalence of E. vermicularis infections presented a tendency towards a rise with age (χ²_trend = 8.373, P < 0.05) and school grade (χ²_trend = 30.274, P < 0.05). Logistic regression analysis identified rural kindergartens and high grades as risk factors, and separate washing of children’s and adults’ cloths, frequent bathing and frequent dinnerware disinfection in kindergartens as protective factors for E. vermicularis infections among children. In addition, there was no significant difference in the detection of E. vermicularis infections among children by using adhesive (73.08%, 76/104) and scotch cellophane-tape anal swab methods (56.73%, 59/104) (χ² = 3.959, P > 0.05). Conclusions The prevalence of E. vermicularis infection is high among children in Fanxian Country, Henan Province. Health education and surveillance of enterobiasis are required to be intensified among children in rural kindergartens and senior grades and their parents and teachers.

·AIM: To observe the clinical efficacy of fumigation treatment of traditional Chinese medicine(Four Yellow Qing Ling Water) for dry eye, and to provide the reference for clinical treatment of dry eye. · METHODS: Totally 82 patients (164 eyes) were randomly divided into two groups from June 2016 to December 2016 in Ophthalmology Department of our hospital. The patients in control group were given artificial tears;the patients in the observation group were given artificial tears and fumigation treatment of traditional Chinese(Four Yellow Qing Ling Water) once a day. After treatment for 14d, the SchirmerⅠtest (SⅠt), break-up time (BUT), cornea fluorescein staining (FL) and clinical efficacy of two groups were compared. ·RESULTS:The efficiency rate of observation group was significantly better than the control group (87. 8% vs 70.7%,P<0.5). The SⅠt and BUT in the observation group were significantly higher than those in the control group (8.43 ± 2.51mm/5min vs 6.38 ± 2.52mm/5min, P<0.05;8.60±2.47s vs 6.35±2.29s, P<0.05); the FL in the observation group (0.84 ± 0.75 vs 1.26 ± 0.84, P<0.05) significantly lower than those in the control group. ·CONCLUSION: The fumigation treatment of traditional Chinese medicine (Four Yellow Qing Ling Water) combined with artificial tears for dry eyes can improve the clinical symptoms of dry eye syndrome.

OBJECTIVETo observe the levels of pulmonary surfactant proteins A and D (SP-A, SP-D) in bronchoalveolar lavage fluid (BALF) of children with pneumonia, and to explore their relationships with clinical characteristics.

METHODSThirty-five children with pneumonia were enrolled in this study. Differential cell counts were obtained by Countstar counting board. The levels of SP-A and SP-D in BALF were detected using ELISA.

RESULTSIn children with pneumonia, SP-D levels were significantly higher than SP-A levels (P<0.001). SP-D levels were negatively correlated with the neutrophil percentage in BALF (r(s)=-0.5255, P<0.01). SP-D levels in BALF in children with increased blood C-reactive protein levels (>8 mg/L) were significantly lower than in those with a normal level of C-reactive protein (P<0.05). Compared with those in children without wheezing, SP-D levels in children with wheezing were significantly lower (P<0.01). There was no correlation between SP-A levels and clinical characteristics.

CONCLUSIONSSP-D levels in BALF are significantly higher than SP-A levels, and have a certain correlation with clinical characteristics in children with pneumonia. As a protective factor, SP-D plays a more important role than SP-A in regulating the immune and inflammatory responses.

Bronchoalveolar Lavage Fluid ; chemistry ; C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant ; Male ; Pneumonia ; metabolism ; Pulmonary Surfactant-Associated Protein A ; analysis ; Pulmonary Surfactant-Associated Protein D ; analysis

Objective To summarize our 5 years experiences of one-stop hybrid procedure (OHP) for the management of congenital heart disease (CHD) in neonates and young children (<2 years old). Methods Clinical data derived from consecutive 152 young children and neonates with CHD underwent OHP between March 2004 to March 2009 were analyzed. Patients were divided into 3 groups: Balloon plasty group (n=72), device closure group (n=43) and collateral arteries occlusion group (n=37). All procedures were image-guided and performed in a specially designed hybrid operation room. Incidence of major adverse cardiovascular events was obtained. Results Patients received successful per-ventricular valvuloplasty or per-aortic balloon angioplasty in balloon plasty group. Two patients in this group with severe right ventricle outflow obstruction received regular open-heart outflow tract rconstruction immediately (n= 1) or selective conventional open-heart operation after discharge (n=1). One neonate with pulmonary atresia with intact ventricular septum died from liver failure 6 month after OHP. In device closure group, device closure was failed in 3 cases (2 with atrial and 1 with ventricular septum defects), 1 young child with vcntricular septum defects died from pneumonia after successful device closure. No device malposition was observed in device closure group during the follow-up. All patients received major collateral arteries occlusion and open-heart correction were discharged without complication. Conclusion OHP could avoid or shorleu the application of cardiopulmonary bypass and reduce the surgical trauma in selected young children with CtiD. Although OHP was feasible and safe, the image outfits, image-guided technology and OHP-related device should be further developed and improved for better procedure outcome.

OBJECTIVETo elucidate the relative factors and prognostic impact for angiographic no-reflow phenomenon during primary percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI).

METHODSA total of 930 patients with AMI who underwent primary PCI were divided into no-reflow group and normal-reflow group. Factors related to no-reflow were analyzed by logistic regression model and major adverse cardiac events (MACE) in-hospital as well as during long-term follow-up was also observed.

RESULTSNo-reflow occurred in 82 out of 930 patients. Admission glucose level (9.8 +/- 4.3 mmol/L vs. 8.5 +/- 3.5 mmol/L, P = 0.001), peak CK-MB value (369.4 +/- 167.8 U/L vs. 282.3 +/- 161.7 U/L, P < 0.01) and percentage of TIMI flow grade 0 on initial angiogram (69.5% vs. 54.5%, P = 0.009) were significantly higher and pre-infarction angina (19.5% vs. 48.1%, P < 0.01) was significantly lower in no-flow patients than normal flow patients. Logistic regression analyses showed that admission glucose level, absence of pre-infarction angina, TIMI flow grade 0 on initial angiogram and pump failure were independent predictors of no reflow. In-hospital MACE (37.8% vs. 11.3%, P < 0.01) and follow-up MACE (37.5% vs. 17.4%, P < 0.01) were also significantly higher in no-flow patients than normal flow patients. Multivariate Cox regression analysis revealed that no-reflow was an independent predictor of long-term cardiac death (relative risk 3.83, 95% confidence interval 1.71 to 5.57).

CONCLUSIONAdmission glucose level, absence of pre-infarction angina, TIMI flow grade 0 on initial angiogram and pump failure were independent risk factors for no-reflow and no-flow is an independent predictor for increased in-hospital and follow-up MACE.

Aged ; Angina Pectoris ; Angioplasty, Balloon, Coronary ; Blood Glucose ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; therapy ; Prognosis ; Treatment Outcome

OBJECTIVEIt has been reported that neuronal apoptosis plays a critical role in pathology of hypoxic-ischemic encephalopathy (HIE). Cytochrome C (CytC) is an important apoptotic protease activating factor. Inosine might have a neuroprotective effect against cerebral ischemia reperfusion injury by inhibiting the neuronal apoptosis and the expression of CytC mRNA in adult rats. This study examined the effects of inosine on neuronal apoptosis and CytC mRNA expression following hypoxic-ischemic brain damage (HIBD) in order to investigate the neuroprotectivity of inosine against cerebral ischemia injury in neonatal rats and the possible mechanism.

METHODSA total of 140 healthy 7-day-old Sprague-Dawley rat pups were randomly assigned into Control (n=40), HIBD (n=50) and Inosine treatment groups (n=50). HIBD rat models were established by ligating the left common carotid artery, followed by 8% O2 hypoxia exposure for 2 hrs in the HIBD and Inosine treatment groups. The Control group was not subjected to hypoxia-ischemia (HI). The Inosine treatment and the HIBD groups were randomly divided into 5 sub-groups sacrificed at 6 and 12 hrs, and 1, 3 and 7 days post- HI (n=10 each). The Control group rats were sacrificed at the corresponding time points (n=8 each). Inosine was administered to the Inosine treatment group by intraperitoneal injection immediately after HIBD at the dosage of 100 mg/kg twice daily for 7 days. TUNEL staining and in situ hybridization method was used to detect neuronal apoptosis and CytC mRNA expression respectively.

RESULTSFew apoptotic cells and CytC mRNA positive cells were found in brain tissues of the Control group. In the HIBD group, the number of apoptotic cells and the CytC mRNA expression in the cortical and hippocampal gyrum CA1 areas increased 6 hrs after HI, peaking at 1 day after HI and then decreased gradually. Until the 7th day, the number of apoptotic cells and the CytC mRNA expression in the cortical and hippocampal gyrum CA1 areas in the HIBD group remained significantly higher than in the Control group. Inosine treatment decreased the apoptotic cells and the CytC mRNA expression in both areas from 6 hrs to 7 days after HI compared with the HIBD group. The linear correlation analysis demonstrated that the number of apoptotic cells was positively correlated to the CytC mRNA expression in neonatal rats with HIBD (r=0.88, P < 0.01) .

CONCLUSIONSInosine can reduce the number of apoptotic cells and down-regulate the expression of CytC mRNA following HIBD in neonatal rats. The decreased number of apoptotic cells was positively correlated to the decreased CytC mRNA expression after inosine treatment, suggesting that inosine offered neuroprotectivity against HIBD possibly through inhibiting the CytC mRNA expression and resulting in a decrease of cell apoptosis.

Animals ; Apoptosis ; drug effects ; Cytochromes c ; genetics ; Hypoxia-Ischemia, Brain ; drug therapy ; metabolism ; pathology ; In Situ Nick-End Labeling ; Inosine ; pharmacology ; therapeutic use ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley