The cell-free plasma DNA (cfpDNA) has been suggested as a useful tumor marker for its quantitative and qualitative tumor-specific alterations that reflect the biological characteristics and the progression and outcomes of tumors.Therefore,it has been used as liquid biopsy to detect cfpDNA in peripheral blood for the diagnosis,monitoring of clinical effects,and prognosis of malignancies

Objective To investigate the growth inhibition and DNA damage of energized fusion protein anti-CD20(Fab)-LDM on B JAB cells in vitro.Methods The binding activity of fusion protein anti-CD20 (Fab)-LDP to B JAB cells was studied by flow cytometry and confocal laser scanning microscopy.MTT assay was used to study the energized fusion protein anti-CD20(Fab)-LDM on cell growth of B JAB cells.Comet assay was employed to detect DNA damage in B JAB cells.The cell growth cycle of BJAB was analyzed by FACS.Results The recombinant fusion protein anti-CD20 (Fab)-LDP possessed an significant target affinity towarded BJAB cells.The energized fusion protein anti-CD20(Fab)-LDM showed obvious inhibition on proliferation,as well as induced potent DNA damage in B JAB cells in vitro compared with lidamycin.B JAB cells treated with energized fusion protein anti-CD20 (Fab)-LDM showed S phase cell cycle.Conclusion The energized fusion protein anti-CD20 (Fab)-LDM could target binding to BJAB cells and significantly inhibit the proliferation of B JAB cells by inducing DNA damage and S phase arrest.

BACKGROUND: Current studies on CD62P have focused mainly on cardiovascular diseases, while only few studies have evaluated the effects of CD62P on the development of sepsis and the association between endothelial cell injury with inflammation and coagulation. This study attended to explore the association between endothelial cell injury with inflammation and coagulation by evaluating the expression of soluble CD62P (s-CD62P) in plasma and its mechanism in patients with sepsis, thus to provide the evidence of effective treatment of sepsis with anti-adhesion therapy targeted CD62P. METHODS: A total of 70 critically ill patients with systemic inflammatory response syndrome (SIRS) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrol ed for a prospective and control study. According to the diagnostic criteria of sepsis/SIRS, the patients were divided into two groups: a sepsis group (n=38) and a SIRS group (n=32). Another 20 healthy volunteers served as a control group. Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure, diabetes and its complications. The demographics of the patients including age, sex, body mass index (BMI), smoking and alcohol addict were compared among the groups. Six mL peripheral blood samples were collected within 24-hour admission in ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P, TNF-α, and hs-CRP. And variables of coagulation function such as platelet (PLT), prothrombin (PT), activated partial thromboplastin time (APTT), D-dimer and antithrombin-III (AT-III) were analyzed during 24 hours after admission to ICU. Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated. Data were expressed as mean±standard deviation and were statistical y analyzed by using SPSS 17.0 statistical software. The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and the Kruskal-Wallis test. The relations between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson's product moment correlation coefficient analysis. Changes were considered as statistically significant if P value was less than 0.05. RESULTS: Compared with the control group and SIRS group, the sepsis group demonstrated significantly higher levels of s-CD62P, TNF-α and highly sensitive C-reactive protein (hs-CRP) (P<0.05). The plasma levels of D-dimer, PT, and APTT in the sepsis and SIRS groups were significantly higher than those in the control group, while the platelet count and the activity of AT-III were obviously lower (P<0.05). In the sepsis group, the plasma levels of hs-CRP and TNF-α were positively correlated with PT, APTT, and D-dimer, and negatively correlated with AT-III and PLT (P<0.05). The plasma levels of s-CD62P were significantly correlated with the plasma levels of TNF-α, hs-CRP, D-dimer, PT, and APTT, whereas they were correlated negatively well with PLT and AT-III (P<0.05). CONCLUSIONS: The concentration of plasma s-CD62P is elevated as a early biomarker in patients with sepsis, and it serves as one of the pathogenic factors responsible for endothelial cell damage. Coagulation and mediators of inflammation promote each other, aggravating the severity of sepsis. Plasma s-CD62P may be an important factor for the development of coagulation and inflammatory reaction.

Objective:To investigate the prevalence of post-traumatic stress disorder(PTSD)symptoms in middle school students after Wenchuan earthquake.Methods:The PTSD Checklist-Civilian version( PCL-C),which included three symptom groups(A,B,and C),was used to assess the PTSD symptoms in 1960 middle school students in the disaster region through self-questionnaire.Results:(1)The general positive rate of PTSD symptoms was 78.3%,and that of B group was the highest(68.9%).(2)The rates of general PTSD and three symptom groups of PTSD were higher in girls,high grade students,minority groups,rural students,injured in earthquake and those who lost family property than in boys,low grade students,the Han nationality,urban students,not injured in earthquake and those who did not lost family property[such as,the general positive rate of PTSD symptoms:girls 82.2%,boys 73.9%,P<0.05].(3)The rates of severe PTSD symptoms in girls and rural students were higher than that in boys and urban students(27.9% vs.19.9%,26.7% vs.21.4%,Ps<0.05).Conclusions:PTSD symptoms are common among middle school students in earthquake region.Targeted measures should be taken to protect middle school students from PTSD in earthquake region.

Objective: To observe the effects of electroacupuncture (EA) on the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment (CI), and to explore the mechanism of EA in improving the learning and memory abilities. Methods: A total of 100 Sprague-Dawley (SD) rats were divided into a normal group (n=10) and a model group (n=90) by random number table method. Rats in the model group were intraperitoneally injected with a small dose of streptozotocin (STZ) to establish the type 2 diabetic models, after being fed with high-fat and high-sugar diet for 1 month. Twenty CI rats were selected from the 50 successful model rats by the Morris water maze (MWM) test and randomly divided into a model group and an EA group according to the blood glucose level and MWM data (n=10). Rats in the EA group received acupuncture at Zusanli (ST 36), Neiting (ST 44) and Yishu (Extra), of which Zusanli (ST 36) and Neiting (ST 44) were stimulated by EA apparatus, 20 min/time, once a day for 6 d a week and 4 consecutive weeks. The rats in the model and the normal groups were fixed without treatment. After 4-week treatment, the random blood glucose level of the rats was measured; the learning and memory abilities of rats were measured by MWM; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptotic cells; Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex. Results: After modeling, the random blood glucose level and the escape latency tested by MWM were significantly increased, and the number of crossing the platform tested by the MWM was decreased in the EA and model groups, and were significantly different from those in the normal group (P<0.05 or P<0.01), while the differences between the model group and the EA group were not statistically significant (all P>0.05). After 4-week treatment, the random glucose level and the escape latency tested by MWM were significantly increased (both P<0.05), and the number of crossing the original platform tested by the MWM was significantly reduced (P<0.01), the protein and gene expressions of Bax and Caspase-3 were significantly increased (all P<0.001), the protein and gene expressions of Bcl-2 were significantly reduced (both P<0.001), and the number of neuron apoptosis was significantly increased (P<0.001) in the model group than in the normal group; the random blood glucose level was significantly reduced (P<0.05), the escape latency tested by MWM was significantly shortened (P<0.05), and the number of crossing the original platform tested by MWM was significantly increased (P<0.05), the protein and gene expressions of Bax and Caspase-3 were significantly reduced (all P<0.001), the protein and gene expressions of Bcl-2 were significantly increased (both P<0.001), and the number of neuron apoptosis was significantly reduced (P<0.001) in the EA group than in the model group. Conclusion: EA can improve the learning and memory damages induced by type 2 diabetic model rats with CI; the action mechanism may be achieved via anti-apoptosis.

In this study, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the changes in mRNAs levels of preproadrenomedullin (ppADM) gene encoding adrenomedullin (ADM) and the essential receptor components of ADM, calcitonin receptor-like receptor (CRLR), and the receptor activity modifying protein 2 and 3 (RAMP2 and RAMP3) in the medulla oblongata, hypothalamus, midbrain, pituitary gland and adrenal gland of the stress-induced hypertensive rats. It was shown that chronic foot-shock and noise stress for 15 consecutive days induced a significant increase in systolic blood pressure (SBP) and unique changes in ppADM and its receptor components mRNAs in all areas studied. As compared with the control group, the level of ppADM mRNA, normalized against a glyceraldehydes-3-phosphate dehydrogenase (GAPDH) control, was up-regulated in the hypothalamus-pituitary-adrenal (HPA) axis, but down-regulated in the medulla oblongata and midbrain (P<0.01 and P<0.05, respectively). The relative amount of CRLR mRNA was higher in the hypothalamus than that in other areas. The level of CRLR mRNA expression was significantly increased in the medulla oblongata of the stress group (P<0.01), but decreased in the midbrain (P<0.01) as well as hypothalamus(P<0.05), as compared with that of the control group. Chronic stress for 15 consecutive days produced an increase in the level of RAMP2 mRNA expression in the medulla oblongata (P<0.01) and a decrease in the adrenal gland (P<0.01), as compared with the control. No significant stress-related changes in RAMP2 mRNA were observed in the midbrain, hypothalamus and pituitary gland. The amount of RAMP3 mRNA was relatively higher in the midbrain and hypothalamus than that in the medulla oblongata, adrenal gland and adrenal gland. Stress-induced hypertensive rats exhibited an increased RAMP3 mRNA expression in the hypothalamus and pituitary gland (P<0.01 and P<0.05, respectively) and a decrease in the adrenal gland and midbrain (P<0.05). No significant stress-related change in RAMP3 mRAN was observed in the medulla oblongata. Taken together, our results indicate that the significant changes in ppADM and its receptor components mRNAs expression in the HPA axis and autonomic centers may be related to the development of the stress-induced hypertension. Nevertheless, the pathophysiological significance of brain-derived ADM and its receptors in stress and blood pressure regulation and their roles in stress-induced hypertension still await further investigation.
Adrenomedullin ; Animals ; Brain Stem ; metabolism ; Hypertension ; etiology ; metabolism ; Hypothalamo-Hypophyseal System ; metabolism ; Male ; Peptides ; genetics ; metabolism ; Pituitary-Adrenal System ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenomedullin ; Receptors, Peptide ; biosynthesis ; genetics ; Stress, Physiological

The purpose of the study was to investigate whether the small fiber-evoked dorsal root reflex (DRR) can be obtained by electrical stimulation of the peripheral nerve in rats. Fifty-one DRRs were recorded from different kinds of fiber filaments in the proximal ends of the cut L(5) dorsal root following electrical stimulation of the sural nerve. According to the kind of afferent fibers in the sural nerve associated with the DRRs from dorsal root, these DRRs were divided into five different types: A(alphabeta) fiber-evoked A(alphabeta).DRR (A(alphabeta)- A(alphabeta).DRR), A(betadelta) - A(delta ).DRR, A(betadelta)-C.DRR, A(alphabetadelta)-C.DRR and C-C.DRR. The results obtained show that the DRR can be obtained from either A-fibers (including A(delta )-fibers) or C-fibers of dorsal root filaments by stimulation of the sural nerve. It is therefore suggested that either A(delta ).DRR or C.DRR can be used as a validity index of presynaptic inhibition of the thin primary afferent terminals for investigation of the modulation mechanisms of peripheral effectors.
Afferent Pathways ; Animals ; Electric Stimulation ; Female ; Male ; Nerve Fibers, Myelinated ; physiology ; Nerve Fibers, Unmyelinated ; physiology ; Rats ; Rats, Sprague-Dawley ; Reflex ; physiology ; Spinal Nerve Roots ; physiology ; Sural Nerve ; physiology

To investigate the correlation between methylation and expression of multidrug resistance (mdr1) gene, restriction endonuclease HpaII combined with competitive PCR technique was used to quantitatively detect the methylation status of two CCGG sites located at -110 and -50 bp (region I and II) up to the transcription start site in mdr1 promoter in 54 AL and 9 MM patients. Semi-quantitative RT-PCR was used to detect the expression level of mdr1 gene. The results showed that inverse correlation between methylation rate of either region or total methylation rate and expression of mdr1 gene was observed. The correlation in the region I (r = -0.64) was closer than that in the region II (r = -0.4). High expression rate of mdr1 ascended significantly in low methylation group (n = 36) (P < 0.001). In comparison with chemotherapy sensitive group (n = 8), the methylation rate in refractory AL patients (n = 16) was lower (P = 0.05) in the region I, P < 0.05 in the region II and total regions. Comparing with the untreated patients (n = 36), the methylation rate in the region I and total methylation rate were lower in the patients with chemotherapy (n = 14) (P < 0.05). The methylation rate in the region II was also decreased after chemotherapy, however, no statistical significance was shown (P > 0.05). Increased mdr1 expression level accompanying with decreased methylation rate after chemotherapy was found, although no significant difference was shown (P = 0.06). It is concluded that the expression level of mdr1 gene was associated with the methylation status of CCGG in -110 and -50 bp upstream to the transcription start site, especially the -110 site. In both the patients treated with chemotherapy and the refractory patients, the methylation level of mdr1 gene decreased relatively. The rising expression of mdr1 gene after chemotherapy was associated with the decrease of methylation level.
DNA Methylation ; Genes, MDR ; Hematologic Neoplasms ; drug therapy ; genetics ; Humans ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the apoptosis of NK cells induced by the erythroleukemia cell line K562 in vitro.

METHODSPrimary NK cells isolated from the peripheral blood of healthy donors by magnetic-activated cell sorting were cultured with stem cell medium containing recombinant human interleukin-2 (rhIL-2). The NK cells and K562 cells were mixed and co-cultured at different E:T ratios for different time lengths. The apoptosis of NK cells and K562 cells were detected using PE-AnnexinV/7-AAD labeling and flow cytometry.

RESULTSThe purity of isolated NK cells reached (93.99∓4.22)%. At the same E: T ratio, the apoptotic rate of NK cells induced by K562 cells increased significantly with time. As the E:T ratio reduced, the apoptotic rate of the NK cells increased and their cytotoxic activity against K562 cells was attenuated.

CONCLUSIONK562 cells can induce the apoptosis of activated NK cells, which is one of the probable mechanisms of immune escape of tumors.

Objective:To classify the symptom characteristics of young and middle-aged peritoneal dialysis patients and to explore the differences of demographic functional status and social function of patients with different symptom categories.Methods:A total of 179 peritoneal dialysis patients from 3 peritoneal dialysis centers in Shanghai were investigated from December 2019 to August 2020 by General Information Questionnaire, Peritoneal Dialysis Symptom Distress Scale. Latent class analysis was used to classify young and middle-aged peritoneal dialysis patients according to symptom characteristics. Multiple Logistic regression was used to explore the differences of demographic and disease characteristics of different categories of patients.Results:Peritoneal dialysis patients could be divided into three potential categories according to symptom characteristics ( P<0.05). According to the conditional probability of each category, they were named "low symptom group" (111 cases,62.0%), "high psychological-moderate physical symptom group" (22 cases, 12.3%), "high symptom group" (46 cases,25.7%). There were differences in working status ( OR=0.029, P<0.01), education level ( OR=152.799, P<0.01), duration ( OR=81.307, P<0.05), diabetic nephropathy ( OR=80.619, P<0.01) and CCI score ( OR=91.188, P<0.01) distribution among different potential categories of young and middle-aged peritoneal dialysis patients. Conclusions:There are three types of potential symptoms in young and middle-aged patients undergoing peritoneal dialysis. In clinical practice, medical staff should focus on the psychological status of young and middle-aged patients with low educational background and early stage of dialysis, and encourage them to return to work; at the same time, they should regularly evaluate the symptom burden of patients with diabetic nephropathy and high complication index peritoneal dialysis patients, in order to provide targeted intervention measures to prevent the progression of the disease.