Objective: To investigate the impact of electrical stimulation numbness to central nervous system in normal people. Methods: Electric stimulator was used to establish low-frequency, high-frequency numbness models with 22 healthy subjects. Scores for numbness and emotional valence for different electrical stimulations were recorded, and data of functional MRI (fMRI) in task state were also collected. The differences of numbness and emotional valence score of subjects under low frequency and high frequency electrical stimulation were compared, respectively. Then brain regions with significant differences in brain activation intensity under different frequencies of electrical stimulation were obtained, and the differences of activation effect values of activated brain regions under different electrical stimulations intensity were compared, respectively. The relationship of numbness and emotional valence scores under high frequency electrical stimulation were analyzed. Results: There were statistical differences of numbness (t=13.18) and valence score (t=10.77) under different electrical stimulations (both P<0.05). There was negative correlation between numbness and valence scores under high-frequency electrical stimulation (r=-0.53, P=0.01). The brain areas with significant differences of activation intensity under different electrical stimulation included left parietal operculum, left middle cingulate gyrus, left temporal gyrus, right parietal operculum, left postcentral gyrus, right central operculum, right posterior insula and left thalamus (all P<0.05). Conclusion: The activated brain regions under different electrical stimulations caused numbness include left parietal operculum, left middle cingulate gyrus, left temporal gyrus, right parietal operculum, left postcentral gyrus, right central operculum, right posterior insula and left thalamus in normal people. Block-designed BOLD-fMRI can be used to detect activity of brain areas associated with numbness.

A recombinant plasmid pET28a-HBcAg-delta n was constructed, in which three mimic B-epitopes of HER family were inserted into the truncated HBc vector. The fusion protein expressed was purified and used to immunize BALB/c mice to induce antibody against the epitopes. Three mimic epitope genes were inserted into the sequences of amino acid residues 78 and 79 of HBcAg by overlap PCR. The PCR product was then cloned into pET28a to construct recombinant expression plasmid which was transformed to E. coli BL21 (DE3) and induced by IPTG. After purification, the fused protein designed HBHE was used to immunize BALB/c mice to detect humoral immunoresponse. The recombinant plasmid was successfully constructed by DNA sequencing analysis. A fusion protein with correct molecular mass was expressed and confirmed by SDS-PAGE. High titre antibody was elicited in the mice immunized with HBHE by indirect ELISA and Western blotting. The HBc particle vector containing three B-epitopes of HER family had been successfully prepared, purified and high titre antibody against HBHE was detected. All these data are helpful in further research of the broad-spectrum anti-tumour effect of combine polypeptide epi-position vaccine of EGFR and HER2.

Objective To investigate the differences of protein and mRNA expression of matrix metalloproteinase 9 (MMP-9) in osteoclasts (OC) and osteoclast-like cells (OLC) obtained by mechanical and inducement methods. Methods Mechanical separation method was used to separate mature osteoclasts from long bones of SD rats aged one-day;and inducement culture method was applied to induce OLC by using RANKL (100 ng/mL) and M-CSF (100 ng/mL) . The protein expression of MMP-9 was measured by immunocytochemistry and mRNA expression of MMP-9 was assayed by in situ hybridization. Results The integral optical density (IOD) and average optical density (AOD) of positive cells in the visual field were higher in the 12-d group of inducement method as compared with the 3 d-group of mechanical method. Conclusions It is suggested that the protein and mRNA expression of MMP-9 in OLC obtained by 12 d inducement method is much high than in OC obtained by 3 d mechanical method. OLC obtained by inducement method can be applied in the study of osteoporosis.

OBJECTIVETo study luteolin-induced non-small cell lung cancer cell line A549 apoptosis and the molecular mechanism for inhibiting its cycle arrest (G2 stage).

METHODMTT assay showed that luteolin had obvious inhibitory effect on A549 and indicated the half inhibition ratio (IC50). Cell cycle and apoptosis were detected by Hoechst 33258 nuclear staining assay, Annexin V-FITC/PI double staining and flow cytometry. Western blotting assay revealed changes in cycle and apoptosis-related proteins induced by luteolin. Possible molecular mechanism was suggested by Western blotting and immunocytochemistry.

RESULTLuteolin had an obvious growth inhibitory effect on A549 cells, with IC50 of 45.2 micromol x L(-1) at 48 h. Flow cytometry showed A549 cells mainly arrested in G2 stage after being treated by luteolin, with low expressions in cyclin A, p-CDC2 and p-Rb. Hoechst 33258 nuclear staining and Annexin V-FITC/PI double staining showed that the luteolin treatment group showed a significant apoptosis rate than the non-treatment group. Western blotting found luteolin can increase phosphorylation of JNK and decrease that of NF-kappaKB (p65). Immunocytochemistry results revealed luteolin can inhibit TNF-alpha-stimulated p65 from nuclear translocation as a transcription factor and thus promoting cell apoptosis.

CONCLUSIONLuteolin can obviously induce apoptosis of human non-small cell lung cancer cell A549 possibly by increasing phosphorylation of JNK to activate mitochondria apoptosis pathway, while inhibiting NF-kappaB from nuclear translocation as a transcription factor.

Annexin A5 ; metabolism ; Apoptosis ; drug effects ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Humans ; Luteolin ; pharmacology ; NF-kappa B ; metabolism

Objective To investigate the clinical value of matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS)in rapid detection of carbapenemase produced by Klebsiella pneumoniae and Pseudomonas aeruginosa.Methods A total of 60 strains of Klebsiella pneumoniae and 80 strains of Pseudomonas aeruginosa isolated from Pizhou People's Hospital affiliated to Xuzhou Medical University from January 2022 to October 2023 were collected,including 30 strains of carbapenem-resistant Klebsiella pneumoniae(CRKP),30 strains of carbapenem-sensitive Klebsiella pneumoniae(CSKP),50 strains of carbapenem-resistant Pseudo-monas aeruginosa(CRPA)and 30 strains of carbapenem-sensitive Pseudomonas aeruginosa(CSPA).Three detection methods were applied,i.e.,modified carbapenem inactivation method(mCIM),colloidal gold immunochromatography and Autof MS 1000 mass spectrometry identification system to evaluate the ability of Autof MS 1000 mass spectrometry identification system in detecting carbape-nase production of Klebsiella pneumoniae and Pseudomonas aeruginosa.Results The results of Autof MS 1000 mass spectrometry iden-tification system were consistent with those of both mCIM and colloidal gold immunochromatography.Carbapenemase was detected in 28 of the 30 CRKP strains,and it was negative in 2 CRKP strains.Carbapenamase was detected in 15 of the 50 CRPA strains and it was negative in 35 CRPA strains.Thirty strains of CSKP and 30 strains of CSPA were all Carbapenemase negative.The coincidence rate of the results of the three methods in the detection for carbapenase was 100％.Conclusion The result of Autof MS 1000 mass spectrome-try identification system has been consistent with those of mCIM and colloidal gold immunochromatography.It not only has the charac-teristics of cost-saving compare with of mCIM method,but also hold the advantages of fast speed and high accuracy of colloidal gold im-munochromatography method.Thus,Autof MS 1000 system can be used for the rapid identification of carbapenemase produced by Kleb-siella pneumoniae and Pseudomonas aeruginosa.

Objective To observe the value of vector flow mapping(VFM)technique for assessing changes of left ventricular diastolic function in ovarian cancer(OC)patients who underwent postoperative chemotherapy.Methods Totally 37 OC patients who received postoperative chemotherapy were prospectively enrolled in chemotherapy group,while 40 healthy adults were taken as controls(control group).Routine echocardiography and VFM were performed for chemotherapy group before chemotherapy,after 3 and 6 cycles of chemotherapy,also for controls at enrollment,and comparison was performed between groups before chemotherapy,as well as among different time points within chemotherapy group,and the correlations of VFM results with hemoglobin and routine echocardiographic results in chemotherapy group were analyzed.Results No significant difference of age,body mass,body surface area(BSA),nor hemoglobin level,routine echocardiographic and VFM results before chemotherapy was found between groups(all P＞0.05).With the process of chemotherapy,hemoglobin level gradually decreased,the isovolumic relaxation period(IR),atrial systole period(AS)intraventricular pressure difference(IVPD)and intraventricular pressure gradient(IVPG)of the left ventricle gradually increased(adjusted P＜0.05),whereas routine echocardiography only showed that the left atrial volume index(LAVI)and the ratio of early mitral inflow velocity and the mean mitral annular early diastolic velocity(E/e')increased after 6 cycles of chemotherapy compared with those pre-chemotherapy(adjusted P＜0.05).In chemotherapy group,VFM results in all diastolic subphases were strongly correlated with hemoglobin levels(|r|=0.718 to 0.836,all P＜0.05),weakly to moderately correlated with LAVI(|r|=0.375 to 0.525,all P＜0.05)and moderately correlated with E/e'(|r|=0.424 to 0.537,all P＜0.05).Conclusion The diastolic function of left ventricle was probably damaged in early stage after postoperative chemotherapy in OC patients.VFM might detect slight changes of early diastolic function of left ventricle more sensitively than routine echocardiography.

Gastrointestinal graft versus host disease is one of the most severe complications after hematopoietic stem cell transplantation, which can occur in patients of any age groups. Its clinical manifestations include nausea, vomit, abdominal pain, diarrhea and the like. Severe gastrointestinal graft versus host disease could directly influence the patients' clinical prognosis and therapeutic efficacy of transplantation. Here we had a review of the research progress on nutritional support and diet management strategies for gastrointestinal graft versus host disease. It is of great clinical significance to form a step-wise nutritional support model to reduce the risk of malnutrition in patients with gastrointestinal graft versus host disease, which would contribute to improving patients' general condition, relieving digestive tract symptoms, and reducing the risk of complications.

Objective To observe the value of gastrointestinal ultrasound contrast agents for evaluating gastric emptying status in patients with superior mesenteric artery syndrome(SMAS).Methods Gastrointestinal ultrasound was prospectively performed in 30 SMAS patients(SMAS group)and 24 healthy subjects(control group).The diameter of proximal duodenum(D),the maximum diameter of duodenum at the angle between superior mesenteric artery and abdominal aorta(d),as well as the degree of duodenum compression(D-d/D)were measured after gastrointestinal contrast agents passing through immediately.Gastric residual volume(GRV)at 5 min(T0),30 min(T1),60 min(T2),90 min(T3)and 120 min(T4)after taking contrast agent were recorded,and the ratio of T1-T4 GRV to T0 GRV(GRV1-GRV4)were calculated and compared between groups.Pearson correlation analysis was performed to explore the correlation of(D-d)/D with GRV1-GRV4 in SMAS patients.Results D,(D-d)/D,GRV1,GRV2,GRV3 and GRV4 in SMAS group were all higher,while d in SMAS group was lower than those in control group(all P＜0.05).(D-d)/D in SMAS patients was moderately and positively correlated with GRV4(r=0.509,P=0.007),but not significantly correlated with GRV1-GRV3(all P＞0.05).Conclusion Gastrointestinal ultrasound contrast agents could be used to real-time dynamically observe gastric emptying status in patients with SMAS and quantitatively analyze GRV.

Objective To develop a risk prediction model for postoperative malnutrition in children with congenital heart disease(CHD)and to verify it both internally and externally.Methods By a convenience sampling method,300 CHD children treated at a tertiary hospital in Shandong Province from January 2018 to December 2021 were selected as a modeling group,and 129 children from January 2022 to June 2023 were selected as a validation group.Data on patient demographics,disease-specific variables,therapeutic interventions,and nursing care parameters were collected.Single factor and logistic regression were employed to construct a risk prediction model for postoperative malnutrition in CHD children,and the nomogram was drawn and its prediction effect was evaluated.Results The incidence of postoperative malnutrition among CHD children was 33.10％.Logistic regression analysis revealed that risk factors for malnutrition in children included birth weight＜2.5 kg,preoperative malnutrition,negative fluid balance 1 week after surgery,and long duration of cardiopulmonary bypass(P＜0.05).The area under the receiver operating characteristic curve of the modeling group was 0.933;the sensitivity was 83.30％;the specificity was 90.90％.The Hosmer-Lemeshow test showed that x2=7.765(P=0.457).The AUC of the validation group was 0.918;the sensitivity was 87.20％;the specificity was 90.00％.The Hosmer-Lemeshow test showed that x2=4.947(P=0.763).Calibration curves for both groups indicated good calibration of the model,and the clinical decision curves demonstrated its practical clinical utility.Conclusion The risk prediction model developed in this study exhibits good predictive ability,which can provide a reference for medical staff to early identify high-risk infants for postoperative malnutrition following CHD surgery and to formulate targeted intervention measures.

Objective: To explore the longitudinal associations between dietary macronutrients and lung function in schoolaged children, so as to provide the nutritional research evidence for promoting children s lung health. Methods: In November 2021, two primary schools located in Chengdu, Sichuan Province were selected from the Southwest China Childhood Nutrition and Growth (SCCNG) cohort by a stratified cluster random sampling method, enrolling a total of 1 112 school aged children aged 8 to 13 years. At baseline, the dietary and sociodemographic characteristics of the children were assessed. One year later, the forced vital capacity (FVC) of the children was measured and converted into Z scores (FVC- Z ), while the vital capacity index (VCI) was also calculated. Generalized linear regression analysis was employed to examine the associations between dietary macronutrients and lung function, considering interactions with gender and age, followed by stratified analysis. Results: After adjusting for confounding factors, the analysis results of the generalized linear regression model showed that the carbohydrate energy ratio was negatively correlated with FVC- Z ( β =-0.02) and VCI ( β =-0.16), while the fat energy ratio showed a positive correlation with FVC- Z ( β =0.03) and VCI ( β =0.23) ( P <0.05). The protein energy ratio was positively correlated with FVC- Z ( β =0.09) and VCI ( β =0.60) specifically in girls ( P <0.05). Additionally, there was an interaction effect of age on the associations between macronutrients and lung function ( P <0.01); in children aged 8-9 and 10-11, the carbohydrate energy supply ratio was negatively correlated with FVC- Z ( β =-0.04, -0.03) and VCI ( β =-0.29, -0.21), and fat energy supply ratio was positively correlated with FVC- Z ( β =0.07, 0.05) and VCI ( β =0.46, 0.32) ( P <0.05). Conclusions There are age and sex differences in the association of dietary macronutrients with lung function, with a low carbohydrate, high fat diet promoting lung function in children. Additionally, protein intake appears to have a positive influence on the lung function of girls. The early school age period may represent a critical window for dietary interventions aimed at promoting lung health.