Objective To prepare anti-apoM monoantibodies with high affinity and high purity, and investigate apoM distribution among human tissues and different groups of people. Methods BALB/c mice were injected intracutaneously with recombinant apoM. After cytomixis, screening and cloning, we established a hybridoma, which grew well and steadily secreted antibodies. The ascites were acquired by injecting BALB/c mice intraperitoneally and anti-apoM monoantibodies were gained using standard techniques. We detected apoM levels in healthy individuals and the patients with coronary heart disease including stable angina (SA) group and acute coronary syndrome (ACS) group using the anti-apoM monoantibodies. ResultsThree anti-apoM monoantibodies were collected and confirmed after subtype identification and block test. The apoM protein were detected in some human cells and human tissues by these three monoantibodies. The concentration of apoM was (11.02 ±1.96) ×10 -3 g/L, (10. 76± 1.32) ×10-3 g/L, (12. 83 ± 2. 28) × 10-3 g/L in SA, ACS and control group respectively. There was significant difference within the three groups (F = 11. 544, P ＜ 0. 05). Comparing apoM concentrations among control group and coronary heart disease groups, it showed that the levels of apoM were lower in coronary heart disease groups than in control group(t =2. 962 and 3. 967,P ＜0. 05). There was no significant difference between two coronary heart disease groups (t = 1. 033, P ＞ 0. 05). Conclusion Anti-apoM monoantibodies are successfully raised and could combine with apoM in human cells and tissues. This lays the foundation for the apoM study in apolipoprotein metabolism.

AIM: To assess the effect of RNAi on suppressing the exogenous reporter gene expression in mammalian neurons,and explore the effect of siRNA quantitation on interference efficiency.METHODS: Exogenous green fluorescent protein(GFP) expression vector was transferred into neuro-2a cells,and then the small interference RNA targeting GFP mRNA(siGFP) synthesized by transcription in vitro at three different concentration was used in this experiment.RESULTS: The results showed that the neuro-2a cells can be transfected efficiently and siGFP can inhibit GFP expression greatly.CONCLUSION: RNAi can be applied into mammalian neurons successfully.The research on siRNA quantitation will provide technique support for studying the gene function of neurons in the future.

Objective To explore the value of contrast-enhanced ultrasound in the diagnosis and classification of traumatic spleen rupture , as compared with enhanced CT. Methods The manifestations of contrast-enhanced ultrasound on surgically or clinically confirmed spleen rupture in 40 patients were retrospectively analyzed. The performance of contrast-enhanced ultrasound on diagnosis and classification was compared with that of enhanced CT. Results For 40 patients with traumatic spleen rupture , the of accuracy of enhanced CT and contrast-enhanced imaging in the diagnosis of traumatic spleen rupture was 97.5%, with no significant statistical differences (chi-square = 0, P = 1). On contrast-enhanced ultrasound examination, 14 patients were diagnosed as true splenic rupture , 9 as subcapsular spleen rupture , and 16 as central splenicrupture, with a accuracy rate of 92.5% (37/40); and accuracy rate for enhanced CT was 90.0% (36/40), there was no significant statistical difference (chi-square = 1.97, P > 0.05). Conclusions Contrast-enhanced and enhanced CT have good consistency in the diagnosis and classification of traumatic spleen rupture. Contrast-enhanced ultrasound can accurately determine the scope and degree of spleen damage , resulting in more accurate classification; and it has values in the diagnosis of traumatic spleen rupture and choice of therapies.

[ ABSTRACT] AIM:To investigate the preventive effects of Clostridium butyricum ( C.butyricum) on the type of pylorus ligated gastric ulcer ( GU) in mice and the underlying mechanisms.METHODS:ICR mice were randomly divided into 4 groups:sham operation group, model group, C.butyricum pretreatment group and omeprazole pretreatment group. Gastric pyloric ligation was adopted to establish GU model in mice.The gastric juice was collected to measure the content of gastric free mucus, the pH of gastric juice and the activity of pepsin.The gastric tissues were collected for routine HE stai-ning to observe the pathological changes.The content of glycogen was detected by PAS staining.The protein expression of Bax and Bcl-2 in the gastric mucosa was also assessed by immunohistochemical staining.RESULTS: The HE and PAS staining showed that the C.butyricum pretreatment obviously attenuated the mucosa lesion induced by ligation.Compared with model group, the pH of gastric juice was significantly raised.The activity of pepsin fell off in C.butyricum group, which was lower than that in omeprazole group.In comparison with model group, the content of gastric free mucus was dra-matically increased and PAS staining showed a significant rise in C.butyricum group, but not in omeprazole group.The protein expression of Bax was decreased and the protein expression of Bcl-2 was upgraded in C.butyricum group than those in model group.CONCLUSION:C.butyricum protects gastric mucosa against the challenge of pylorus ligation in mice and its mechanism may be related to inhibiting gastric acid secretion and the activation of pepsin, increasing the production of gastric free mucus, strengthening the expression of bcl-2 gene and inhibiting the expression of bax gene.

Objective To investigate the sleep quality of pacemaker implanted patients,to analyze the relationships among neuroticism,positive psychological capital and sleep quality,and to examine the potential mediating effect of positive psychological capital on the relationship between neuroticism and sleep quality. Methods Using convenience sampling, a total of 178 pacemaker implanted patients were interviewed and analyzed in two tertiary hospitals of Shandong province. They were assessed with Big Five Inventory,The Positive Psycap Questionnaireand Pittsburgh Sleep Quality Indexfor the evaluation of neuroticism,positive psychological capital and sleep quality.The Process macro was used to test the mediating effect. Results The mean score of PSQI in patients with pacemaker implantation was (8.48± 4.50), and 71.3％of them had sleep problems.The scores of neuroticism had positively correlated with the PSQI scores(r=0.31, P<0.01), the scores of positive psychological capital had negatively correlated with the PSQI scores(r=- 0.39, P<0.01).The confidence interval from Bootstrap output indicated positive psychological capital played a mediating role between neuroticism and sleep quality (95％CI:0.06-0.23), mediation effect was 0.14, the percentage of mediation effect in total effect was 43.75％. Conclusions The relation of positive psychological capital to neuroticism and sleep quality among pacemaker implanted patients are intimate.Positive psychological capital could mediate the relationship between neuroticism and sleep quality. It is suggested that nurses can improve the patient′s sleep by enhancing the intervention of positive psychological capital according to the personality characteristics of patients.

Cholangiocarcinoma is a highly heterogeneous tumor with an insidious onset,severe conditions,a high degree of malignancy,and an extremely poor prognosis.Glucose is a major energy source for the proliferation and metastasis of cholangiocarcinoma,and the glucose metabolism pathway of cholangiocarcinoma cells will be re-edited in the process of rapid proliferation to produce a large amount of energy for their own needs.Traditional Chinese medicine has unique advantages in the treatment of cholangiocarcinoma,and studies have shown that the active components of traditional Chinese medicine can inhibit the development and progression of cholangiocarcinoma by regulating glucose metabolism.This article reviews the characteristics of glucose metabolism in cholangiocarcinoma and the role of the active components of traditional Chinese medicine in regulating glucose metabolism against cholangiocarcinoma,in order to provide new ideas for the treatment of cholangiocarcinoma.

Objective:To investigate the effect of instantaneous flow rate on the consistency of diagnostic accuracy of severe degenerative mitral regurgitation (DMR) using proximal isovelocity surface area (PISA).Methods:From June 2019 to June 2021, 75 patients with DMR who underwent echocardiography in Department of Echocardiography of Zhongshan Hospital, Fudan University were prospectively enrolled. The instantaneous flow rate of DMR during the systolic phase was calculated using M-mode PISA(PISA M-mode), and a time-integrated curve was plotted. Regurgitant volume (RVol) and effective regurgitant orifice area (EROA) were calculated by traditional PISA (PISA max), pair PISA (PISA pair), and PISA M-mode, respectively. RVol acquired from cardiac magnetic resonance (CMR) volumetric method in 22 patients of the enrolled patients. The correlation and consistency of RVol acquired between the three PISA methods and CMR were compared. Agreement of diagnostic accuracy of severe mitral regurgitation (sMR) acquired between the three PISA methods and multi-parameter algorithm by American Society of Echocardiography (ASE) was analyzed using Cohen′s Kappa analysis. Results:The curve of instantaneous flow rate of DMR showed unimodal pattern with the peak at mid-late systolic phase. The correlation of RVol acquired between PISA methods and CMR was moderate for PISA max and PISA pair ( r=0.77, 0.80, both P<0.001), whereas PISA M-mode presented strong correlation with CMR ( r=0.87, P<0.001). RVol acquired from PISA max was larger than that of CMR[(69.1±37.1) ml vs (49.0±29.0)ml, P=0.002]. Both PISA max and PISA pair were shown moderate agreement of diagnostic accuracy of sMR with ASE multi-parameters algorithm (RVol: κ=0.496, 0.525, both P<0.001; EROA: κ=0.570, 0.578, both P<0.001), while PISA M-mode presented strong agreement (RVol: κ=0.867 and EROA: κ=0.802, both P<0.001). Conclusions:Based on the unimodal pattern of instantaneous flow rate in patients with DMR, PISA max may significantly overestimate RVol, exposing a significant proportion of patients with DMR to unnecessary MR surgery. PISA M-mode presents better correlation and consistency with CMR on the quantification of RVol compared with PISA max and PISA pair, and may improve the diagnostic accuracy of quantification of sMR using PISA.

[Objective]To establish the reference intervals of thyroid hormones in pregnant women from Sanming city and compare them with those from other areas in China.[Methods]The study recruited 605 pregnant women and 229 non-pregnant healthy women who visited Sanming First Hospital between March 29 and June 28,2023.Blood samples were se-quentially collected from the participants to determine the serum levels of thyroid stimulating hormone(TSH),free triiodo-thyronine(FT3)and free thyroxine(FT4).The reference intervals of thyroid hormones were established by using a percen-tile range of P2.5 to P97.5 and their changes in different periods of pregnancy were analyzed and compared with those from oth-er areas in China.[Results]There were significant differences in levels of TSH,FT4 and FT3 at different periods of pregnan-cy(all P<0.05).Compared with non-pregnant women,women in first trimester had significantly decreased levels of TSH and FT4,but similar level of FT3.Since the second trimester,TSH level displayed rising tendency,while FT3 and FT4 lev-els showed gradual decrease.The reference intervals of thyroid hormones in the first,second and third trimester of pregnan-cy in Sanming city were TSH(0.068,2.943)μU/mL,FT3(4.302,6.888)pmol/L,FT4(8.240,14.719)pmol/L;TSH(0.419,3.274)μU/mL,FT3(4.074,6.629)pmol/L,FT4(6.726,11.980)pmol/L;TSH(0.422,3.570)μU/mL,FT3(3.741,5.850)pmol/L,FT4(6.103,10.347)pmol/L,respectively.Significant differences were found in the reference in-tervals of thyroid hormones during pregnancy among different areas in China.[Conclusions]The levels of TSH,FT3,FT4 during pregnancy are different from those during non-pregnancy,and also significantly differ in different periods of preg-nancy.Reference intervals of thyroid hormones in pregnant women are affected by various factors such as geographic loca-tion,ethnicity and laboratory test method,etc.Therefore,establishing the population-specific reference intervals of thy-roid hormones will benefit for the diagnosis and treatment of thyroid disease in pregnant women from Sanming city.

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β₂m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a Superdex^TM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals ; Capripoxvirus ; Epitopes, T-Lymphocyte/genetics* ; Peptides/genetics* ; Poxviridae Infections ; Sheep ; Sheep Diseases