Objective To establish an accurate sorting method with less interference to the cell activity for isolating CD 3+CD4+T lymphocytes in human intestinal tissue .Methods Firstly the digestive enzymes were adopted to decompose the human intestinal tissue ,and then the cell suspension was obtained by the filtration of 40 μm cell strainer .Mononuclear cells were collected by using continuous density Percoll gradient centrifugation technique ,then the flow cytometry was adopted to sort CD3+ CD4+ T lympho-cytes in mononuclear cells .The survival rate ,cell purity and morphological observation of the sorted cells were adopted to conduct the evaluation on the sorting method .Results The digestive enzymes well decomposed human intestinal tissue into cell suspension , the purity of CD3+CD4+ T lymphocytes by using continuous density Percoll gradient centrifugation was (43 .9 ± 7 .3)% ,while the purity of CD3+CD4+ T lymphocytes was(96 .9 ± 1 .2)% after fluorescence-activated cell sorting ,the difference between them was statistically significant(P<0 .01) .The survival rate of sorted CD3+CD4+ T lymphocytes was(97 .8 ± 1 .6)% with intact cell shape . Conclusion The Percoll gradient centrifugation technique combined with the fluorescence-activated cell sorting not only collects highly pure human intestinal CD3+CD4+ T lymphocytes with little influence on the cell activity and shape ,but aslo may be carried out under sterile conditions ,the sorted cells can continue to be used in the other functional researches .

Objective:To study the relationship between genetic polymorphisms of GSTM1 GSTT1 and the susceptibility of laryngeal and hypopharyngeal carcinomas(LHC).Method:The GSTM1 an GSTT1 genotypes were determined by multiplex PCR analysis in 76 LHC patients and 76 population controls.The association be tween the genotypes and LHC risk was measured by odds ratios(ORs)and 95% confidence intervals(95%Cls).Resuit:The frequency of GSTM1 null genotype was 59.2% in the LHC patients and 42.1% in controls(OR=1.935,95%CI=1.069-3.510),the difference was significant(P＜0.01).The frequency of GSTT1 null genotype was 57.9% in the LHC patients and 51.3% in controls.The difference was not significant(P＞0.05).In smokers,the risk of the LHC increased in subjects of GSTM1 null genotype(OR=5.545,95%CI=2.158-13.528).Conclusion:GSTM1 polymorphisms are associated with susceptibility to the LHC.It has the synergistic effects with smoking in the development of the LHC.GSTT1 genotypes might have no association with risk of the LHC in urban Linyi.

OBJECTIVE: To study the relationship between genetic polymorphisms of GSTM1 GSTT1 and the susceptibility of laryngeal and hypopharyngeal carcinomas (LHC). METHOD: The GSTM1 and GSTT1 genotypes were determined by multiplex PCR analysis in 76 LHC patients and 76 population controls. The association between the genotypes and LHC risk was measured by odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULT: The frequency of GSTM1 null genotype was 59.2% in the LHC patients and 42.1% in controls (OR=1.935, 95% CI=1.069-3.510), the difference was significant (P<0.01). The frequency of GSTT1 null genotype was 57.9% in the LHC patients and 51.3% in controls. The difference was not significant (P>0.05). In smokers, the risk of the LHC increased in subjects of GSTM1 null genotype (OR=5.545, 95% CI=2.158-13.528). CONCLUSION GSTM1 polymorphisms are associated with susceptibility to the LHC. It has the synergistic effects with smoking in the development of the LHC. GSTT1 genotypes might have no association with risk of the LHC in urban Linyi.
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; Disease Susceptibility ; Female ; Gene Frequency ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Hypopharyngeal Neoplasms ; genetics ; Laryngeal Neoplasms ; genetics ; Male ; Polymorphism, Genetic

Objective To evaluate the effects of continued grateful emotion after discharge on the long-term positive experience and care burden of spouses of patients with brain tumors.Methods Totally 114 patients with brain tumor were selected as the research objects from October 2015 to October 2016. Random number table was used and all patients were randomly divided into the control group (n=56) and the observation group (n=58). Patients in the control group received routine health education and guidance, and two emotional emotional interventions were given to patients during the hospital; while patients in the observation group received increased emotional interventions according to the care burden of spouses of patients. The positive experience and care burden of two groups of spouses were evaluated by Caregiver Positive Experience Scale (PAC) and Caregiver's Burden Scale (CBI) 2 months, 3 months, and 6 months after discharge.Results There was no significant difference in positive experience and care burden of spouses between two groups (P>0.05); 3 months and 6 months after discharge, the scores of positive experience and care burden of spouses in the observation group were [(36.76±4.13), (36.71±4.21)]and [(27.07±4.62), (27.62±4.47)], and they were significantly better than those of the control group [(54.5±3.25),(25.36±3.17)and (54.75±6.27), (54.01±6.21)], the differences were statistically significant (t=7.847-8.587,P<0.05).Conclusions Emotional intervention during hospitalization can effectively improve the positive experience and care burden of spouses of patients. After discharge, emotional intervention can provide outpatient nursing care for the spouses of patients. There is a better long-term effect in improving the positive experience and care burden of spouses of brain tumor patients.

Objective:To evaluate the association between preoperative serum β 2-microglobulin (β 2MG) concentrations and postoperative delirium (POD) in elderly patients. Methods:The study selected patients who underwent knee or hip arthroplasty under spinal-epidural anesthesia on an elective basis at Qingdao Municipal Hospital from May 2021 to November 2022. The patients were divided into a POD group and a non-POD group based on the occurrence of POD. The study was conducted as part of the Perioperative Neurocognitive Impairment and Biomarkers Lifestyle Cohort, which was a nested case-control study. The study collected baseline data from two groups of patients and analyzed the differences between them. Logistic regression was used to identify the risk factors for POD. The stability of the regression model was tested using sensitivity analysis. The mediation model was used to examine whether cerebrospinal fluid (CSF) biomarkers mediated the relationship between β 2MG and POD. The receiver operating characteristic curve was drawn and the area under the curve was calculated to evaluate the accuracy of preoperative β 2MG concentrations and CSF biomarker concentration in predicting POD. Results:There were 57 cases in POD group and 449 cases in non-POD group. The results of logistic regression analysis showed that the increased β 2MG and CSF total tau protein (t-tau) concentrations were risk factors for POD, and the increased CSF β-amyloid 42 concentration was a protective factor for POD after adjustment for multiple confounders such as age, gender, education, Mini-Mental State Examination, history of hypertension and infusion volume ( P<0.05). The results of mediation analysis showed that the serum β 2MG′s effect on POD was partly mediated by t-tau (18.1%). The results of the receiver operating characteristic curve showed that the area under the curve of the β 2MG concentration combined with the CSF biomarker concentration was 0.742. Conclusions:Elevated preoperative serum β 2MG concentration is a risk factor for POD in elderly patients, and the relationship may be partly mediated by CSF t-tau.

Objective:To evaluate the relationship between physical activity (PA) and postoperative delirium (POD) in elderly patients undergoing knee or hip arthroplasty.Methods:The study was conducted as part of the Perioperative Neurocognitive Impairment and Biomarkers Lifestyle Cohort, which was a nested case-control study. Medical records from elderly patients undergoing elective knee or hip arthroplasty under spinal-epidural anesthesia at Qingdao Municipal Hospital from August 2022 to August 2023 were collected. The patients were divided into a POD group ( n=89) and a non-POD group ( n=221) based on the occurrence of POD. Peripheral blood samples were collected before surgery, and the cerebrospinal fluid (CSF) 2 ml was extracted after successful puncture under spinal-epidural anesthesia for determination of the concentrations of amyloid-β 42 (Aβ 42), total tau protein (t-tau), and phosphorylated tau protein (p-tau) by enzyme-linked immunosorbent assay. Logistic regression was used to analyze the influencing factors of POD, and the mediation analysis was conducted to examine the mediating role of CSF biomarker in the relationship between PA and POD. Results:Logistic regression analysis showed that the increased concentration of CSF biomarkers Aβ 42 ( OR=0.997, P=0.006), elevated ratio of Aβ 42/t-tau ( OR=0.642, P=0.003), elevated ratio of Aβ 42/p-tau ratio ( OR=0.872, P=0.001) and PA ( OR=0.374, P=0.001) were protective factors for POD, while the elevated concentrations of t-tau ( OR=1.006, P=0.001) and p-tau ( OR=1.030, P=0.011) were risk factors for POD after adjusting for multi-confounders such as hypertension, diabetes, history of drinking, years of education and Mini-Mental State Examination score. The results of the mediation analysis showed that Aβ 42 (20%), t-tau (16%), Aβ 42/t-tau (23%) and Aβ 42/p-tau (28%) played mediating roles in the relationship between PA and POD. Conclusions:PA is a protective factor for POD in elderly patients undergoing knee or hip arthroplasty and CSF biomarkers may play a mediating role in the relationship between PA and POD.

Objective:To evaluate the relationship between the preoperative sleep quality and postoperative delirium (POD) in elderly patients.Methods:Three hundred and eighty-nine patients of either sex, aged 65-90 yr, with American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, who underwent elective knee/hip arthroplasty under combined spinal-epidural anesthesia in Qingdao Municipal Hospital from May 2022 to October 2023, were selected. The Pittsburgh Sleep Quality Index scale was applied at 1 day before surgery to assess the sleep quality within 1 month before surgery. The cerebrospinal fluid (CSF) specimens 2 ml were withdrawn from the subarachnoid space before anesthesia for determination of the concentrations of biomarkers (β-amyloid), total tau protein [t-tau], and phosphorylated tau protein [p-tau]) by enzyme-linked immunosorbent assay. Patients were divided into POD group and non-POD group based on whether delirium occurred within 7 days after surgery. Logistic regression was used to identify the risk factors for POD, and the sensitivity analysis was used to test the stability of the regression models. The mediation model was used to examine whether CSF biomarkers mediated the link between sleep quality and POD. The accuracy of preoperative sleep quality and CSF biomarker concentrations in predicting the occurrence of POD was evaluated by plotting the receiver operating characteristic curve and calculating the area under the curve.Results:Two hundred and seventy-nine patients were finally included, with 33 patients in POD group and 246 patients in non-POD group. The results of logistic regression analysis showed that shorter sleep duration, use of hypnotics, daytime dysfunction within 1 month before surgery, elevated t-tau concentrations in CSF, and elevated p-tau concentrations in CSF were risk factors for POD after adjusting for multiple confounding factors such as age, sex, body mass index, years of education, Mini-Mental State Examination score, and VAS score. The results of mediating effect showed that the effect of use of hypnotics and daytime dysfunction within 1 month before surgery on POD was partially mediated by p-tau concentrations in CSF (>10%). The area under the curve of combination of daytime dysfunction, sleep duration, use of hypnotics within 1 month before surgery, and CSF biomarker concentrations in predicting POD was 0.979.Conclusions:Shortened sleep duration, use of hypnotic, and daytime dysfunction with 1 month before surgery are risk factors for POD in elderly patients, and the association between use of hypnotics and daytime dysfunction and POD is partially mediated by p-tau in CSF.