Adoptive cellular immunotherapy (ACI) achieves the elimination and control of tumor by mobilizing the body's immune function.It also has targeted efficacy and mild untoward effects.Cytokineinduced killer cells and tumor-infiltrating lymphocytes have been widely used in clinic and have obtained preliminary efficacy.With the development and clinical application of the specific gene transfer of T cell,it will further increase the efficacy of immunotherapy.At present,improving cell culture technology and cell function and using with other treatment are the key links to improve the efficacy of ACI.

Objective:Human adipose-derived stem cells (hASCs)are a highly attractive source in bone tissue engineering.To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs ) in vitro,and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vi-vo.Methods:The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OCpro-Luc-Puro construct.Then,the OCpro-Luc-Puro construct together with three assistant vectors:pM-DLg/pRRE,pRSV-REV,and pVSVG,were transiently transfected into HEK293T cells followed by viral supernatants collection,filtration and concentration.Next,the hASCs stably expressing luciferase repor-ter gene driven by osteocalcin promotor were created with the lentivirus carrying OCpro-Luc-Puro cassette under puromycin selection.The OCpro-Luc-hASCs were then cultured in the absence or presence of osteo-genic differentiation medium.On the 7th and 1 4th days,after osteogenic induction,cellular extracts were collected and analyzed by luciferase reporter assay.Meanwhile,alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR)analysis of osteogenic associated genes osteo-calcin (OC),runt-related transcription factor 2 (Runx2)and alkaline phosphatase (ALP)were used to assess the osteogenic differentiation ability of OCpro-Luc-hASCs.Results:OCpro-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated.On the 7th and 1 4th days after osteogenic induction,the luciferase activity of the cellular extracts from OCpro-Luc-hASCs was dramatically increased.Consistently, the extracellular matrix mineralization,as shown by Alizarin red S (ARS)staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC,Runx2 and ALP, although to variable extent,was observed upon osteogenic differentiation.These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OCpro-Luc-hASCs.Conclusion:We established a luciferase reporter system that could be used to rapidly,quantitatively and specifically determine osteo-genic differentiation ability of hASCs.Comparing with the traditional time-consuming methods,the system we generated here was highly effective.This system not only can be used to examine ostogenic differentia-tion of hASCs in a high throughput manner,but also provides a way to monitor ostogenic differentiation of cells in vivo.

Graphene oxide (GO) modified stationary phase for open tubular capillary (3 m × 25 μm i. d. ) of liquid chromatography ( OT-CLC) was fabricated by coating GO sheets onto poly ( vinylbenzyl chloride-divinylbenzene) porous composite layer via covalent coupling, which was prepared by in-situ polymerization. Scanning electron microscopy ( SEM), Raman spectroscopy and transmission electron microscopy ( TEM) were used to characterize the structure of the stationary phase. The results demonstrated that poly(vinylbenzyl chloride-divinylbenzene) porous composite layer had decentralized globular structure, and the GO sheets covered on the porous layer homogenously. The phase ratio and sample capacity were greatly improved due to the spherical polymer layer and the coverage of GO. Thus, alkyl benzenes, neutral polycyclic aromatic hydrocarbons, acidic and basic compounds could be well separated by using acetonitrile-water as eluent, while four nucleobases were completely separated by using acetonitrile-0. 02 mol/ L ammonium acetate as mobile phase. The run-to-run, day-to-day, and column-to-column reproducibilities were evaluated by calculating the relative standard deviations (RSDs, n =6) of the retention time of o-phenylenediamine, aniline and 2, 4, 6-trifluoroaniline, respectively. These RSD values were all in the range of 0. 3% -2. 0% .

Objective To explore the effect of mental nursing intervention on gastric cancer with liver metastasis patients treated with TACE sequential chemotherapy.Methods Sixty four gastric cancer patients with liver metastasis,no hopes of accepting operations,were included.All patients were divided into control group (32 cases) and observation group (32 cases).The former received routine nursing care and the later received extra mental nursing intervention.The anxiety,depression and quality of life in both groups were evaluated and compared.Results After extra mental nursing intervention,the SDS and SAS scores of the observation group [(41.6 ± 1.8) points and (41.6 ± 2.3) points] were significantly lower than those of the control group [(50.9 ± 1.9) points and (53.0 ± 3.5) points,P =0.040,P =0.036,respectively].The quality of life scores,including the physiological function,body pain,general self rating,vitality,social function,emotional function and mental health score were (45.26 ± 38.26)points vs (35.17 ± 33.65) points (P =0.044),(71.98 ± 17.36) points vs (63.01 ± 18.41) points (P =0.039),(55.59 ± 12.78) points vs (49.67 ± 12.43) points (P =0.048),(74.82 ± 14.23) points vs (55.61 ± 10.58) points (P =0.027),(75.47 ± 12.15) points vs (57.22 ± 14.78) points (P =0.031),(70.24 ± 22.00) points vs (45.92 ± 34.76) points (P =0.004),(78.31 ± 19.11) points vs (67.23 ± 11.96) points (P =0.037),respectively,the scores in the observation group were significantly higher than those in the control group (all P ＜ 0.05).Conclusion The extra mental nursing intervention on gastric cancer with liver metastasis patients,treated with TACE sequential chemotherapy,can not reduce the patients'negative mood,improve the patients'compliance with chemotherapy and the confidence to treatment,but improve the quality of patients'life effectively.

Objective:To investigate the influence and mechanism of Portulaca extracts on Tumor Necrosis Factor -α(TNF-α), Interleukin-4(IL-4) in skins of rats with atopic eczema (AE).Methods:40 SD rats were randomly divided into normal control group , model group,Cortex Phellodendri group and Porulaca group.2,4-dinitrochlorobenzene was used on the right back of rat for making atopic eczema.21 days later , all animals were sacrificed , compared the weight of the same size of the eczema and healthy skin.The levels of TNF-αand IL-4 in rats skins were detected by immunohistochemical method.Results:The right back of rats in model group appeared obvious erythema ,erosion and leakage ,which meant the modeling succeeded.Compared with model group ,the swelling degree of Cortex Phellodendri group and Purslane group were significantly decreased ( P<0.01 ) ,compared with Cortex Phellodendri group ,the swelling degree of Purslane group were less than them ( P<0.05 ).The levels of TNF-α, in rats skins of normal control group , model group,Cortex Phellodendri group and Portulaca group were (6 652.66±1 190.94 ),(19 927.10±5 494.21 ), (7 515.13±877.66 ), (6 809.93±1 385.54).The levels of IL-4 were (5 378.44±1 685.01),(26 334.89±3 993.48),(7 814.84±1 751.38),(8 246.57± 975.08 ).Compared with model group ,the levels of TNF-α,IL-4 in rats skins of Cortex Phellodendri group and Purslane group were sig-nificantly decreased(P<0.01).Compared with positive drug group,the levels of TNF-α,IL-4 in rats skins of Purslane group were a little decreased(P<0.05).Conclusion:Purslane extracts has effect to treat atopic eczema ,its possible mechanism is by erducing the proinflammatory factor ,and moderating the anti-inflammatory factor and the ratio between them.

Objective:To explore the application effect of dyadic intervention scheme based on dyadic disease management theory and Information, Knowledge, Attitude, and Practice model in the discharge preparation of elderly stroke patients and family caregivers.Methods:The 92 pairs of elderly stroke patients and their caregivers hospitalized in the Department of Neurology in People′s Hospital of Zhengzhou University were conveniently selected. The non synchronous control method quasi experimental research was adopted. Totally 46 pairs of subjects who met the criteria for admission and discharge from May to July 2022 were set as the control group, and routine nursing was carried out; from August to October 2022, 46 pairs of subjects who met the criteria for admission and emission were set as the observation group to implement the dyadic intervention program. The scores of discharge readiness, self-efficacy and unplanned readmission rate of patients between the two groups were compared, and the scores of caregiver readiness, self-efficacy and caregiver stress between the two groups were compared.Results:Finally, 85 pairs of subjects completed the study, with 42 pairs in the control group and 43 pairs in the observation group. On discharge day, the total scores of discharge readiness and caregiver readiness in the observation group were (95.19 ± 4.47), (23.02 ± 2.20) points, respectively, which were higher than those in the control group (85.71 ± 5.31), (19.57 ± 1.65) points, with statistically significant differences ( t=8.91,8.16, both P<0.01); the self-efficacy levels of patients in the observation group at discharge and one month after discharge, as well as those of caregivers at discharge and one month after discharge were (73.86 ± 4.87), (75.91 ± 4.51), (75.67 ± 4.99), (79.21 ± 4.90) points, respectively, higher than those in the control group (71.62 ± 5.19), (73.33 ± 4.91), (73.48 ± 4.24), (75.48 ± 4.24) points, with statistically significant differences ( t values were from 2.05 to 3.75, all P<0.05); the pressure levels of caregivers in the observation group at discharge and one month after discharge were (7.51 ± 2.48), (6.28 ± 1.99) points, respectively, lower than those in the control group (8.76 ± 2.55), (7.45 ± 2.36) points, with statistically significant differences ( t=-2.29, -2.48, both P<0.05); the unplanned readmission rate of patients in the observation group one month after discharge was 7.0% (3/43), lower than the control group′s 23.8% (10/42), with statistically significant difference ( χ2=4.65, P<0.05). Conclusions:The implementation of dyadic intervention on elderly stroke patients and caregivers can make their discharge preparation process more adequate, thus reducing the caregiver′s care pressure, reducing the unplanned readmission rate of patients, and improving their health outcomes.

Objective: To examine the associations of pet ownership and daily outdoor activity duration with depressive symptoms in middle school students, so as to provide evidence for targeted prevention strategies of depressive symptoms in middle school students. Methods: Using a stratified cluster random sampling method, 83 601 middle and high school students from 103 districts and counties in Inner Mongolia Autonomous Region were selected in 2024. A questionnaire survey was conducted to collect demographic data, household pet ownership, outdoor activity duration, and depressive symptoms of the research subjects. The comparison of reporting rates of depressive symptoms among different groups of middle school students was conducted using a χ 2 test. The association between pet ownership and outdoor activity duration and depressive symptoms among middle school students was evaluated using a Logistic regression model, and stratified analysis was conducted among different genders and regions to control for potential confounding factors and evaluate the stability of the association. Results: The prevalence of depressive symptoms among middleschool students in Inner Mongolia was 17.2%. Significant differences in depressive symptom reporting rates were observed across sex, grade, ethnicity, surveillance site, parental education, menarche/spermarche status, boarding status, smoking and alcohol use, daily breakfast consumption, school bullying, continuous 30 minute headphone use in a noisy environment, and often use the Internet ( χ 2=8.07-2 672.57, all P <0.01). Both pet ownership ( OR =0.78, 95% CI =0.75-0.81) and ≥2 h/d of outdoor activity( OR =0.81, 95% CI =0.78-0.84) were inversely associated with depressive symptoms;compared to the without owning pets and < 2 h of outdoor activity daily group, students who both owned pets and engaged in ≥2 h of outdoor activity daily had an even lower risk ( OR =0.83, 95% CI =0.78-0.87)(all P <0.05). Conclusion Pet ownership and increased daily outdoor activity duration may help mitigate depressive symptoms among middle school students.

Objective To construct models of viral transfection and hypoxia/reoxygenation induced cellular injury in adult mouse cardiomyocytes(AMCMs)isolated using a non-Langendorff method.Methods AMCMs were isolated,extracted,sedimented,and plated using a non-Langendorff method.The morphology and survival rate of the isolated cells were evaluated 2,24,48 and 72 h after plating,and their integrity was observed by immunofluorescence staining for α-actinin.The isolated AMCMs were infected with adenoviruses carrying an RFP-expressing vector and fluorescence images were obtained at 36 and 48 h post-infection and used to calculate transfection efficiency.The cells were cultured under hypoxic conditions for 45 min,reoxygenated for 24 h,and then stained with propidium iodide(PI)to verify establishment of the hypoxia/reoxygenation injury model.Results The survival rates of AMCMs at 2,24 and 48 h after plating were comparable,but survival was significantly reduced at 72 h.The integrity of the AMCMs was good and＞80%of the cells were transfected with adenovirus at 48 h.After hypoxia/reoxygenation treatment,42%of cells were stained by PI,suggesting successful establishment of the AMCM injury model.Conclusions In this study,we developed a non-Langendorff method for the fast and easy isolation of AMCMs with high cell viability.The isolated cells can be efficiently infected with adenovirus and respond to hypoxia/reoxygenation injury.These findings provide a systematic method for isolating AMCMs and for applying gene modification and hypoxia/reoxygenation injury in these cells.

Objective To determine the risk factors associated with early allograft dysfunction (EAD) after liver transplantation. Methods We retrospectively analyzed the records of 138 patients who underwent liver transplantation from January 2006 to October 2016 in our department. Transplant recipients were divided into two groups:those who met the diagnostic criteria of EAD (EAD group) and those who did not (non-EAD group). We compared blood flow into the transplanted livers and other clinical features between the two groups using univariate and multivariate analysis. Results Intraoperative portal vein flow (PVF) maximum was significantly different between the two groups. Multivariate analysis revealed that intraoperative PVF maximum ＜1 600 mL/min was the only independent risk factor for the occurrence of EAD after liver transplantation in this cohort. Conclusion Intraoperative PVF maximum ＜1 600 mL/min is an independent risk factor for the occurrence of EAD after liver transplantation. Measuring intraoperative blood flow into the transplanted liver in liver transplant recipients may help identify patients at risk for developing EAD.

The development of longitudinal cohorts has made the identification and surveillance of multiple biological markers and behavioral factors which influence disease course or health status become possible. However, traditional statistical methods typically use univariate longitudinal data for research, failing to fully exploit the information from multivariate longitudinal data. The group-based multi-trajectory model (GBMTM) emerged as a method to study the developmental trajectory of multivariate data in recent years. GBMTM has distinct advantages in analyzing multivariate longitudinal data by identifying potential subgroups of populations following similar trajectories by multiple indicators that influence the outcome of interest. In this study, we introduced the application of GBMTM by explaining the fundamental principles and using the data from a health management study in the elderly by using smart wearing equipment to investigate the relationship between multiple life-related variables and hypertension to promote the wider use of GBMTM in longitudinal cohort studies.