Objective To analyze the clinical diagnosis and treatment strategies of liver-localized posttransplantation lymphoproliferative disease (LL-PTLD).Methods Six cases of LL-PTLD from more than 3000 cases of liver transplant recipients from July 2003 to July 2011 were retrospectively analyzed.Other six cases of LL-PTLD were retrieved through Pubmed and Wanfang.The diagnosis and treatment of 12 cases of LL-PTLD were summarized and analyzed.Results All patients with LL-PTLD were diagnosed pathologically.The incidence of LL-PTLD was 0.2％ (6/3000).Among 12 patients,immunosuppressant and anti-EB virus treatment was reduced or withdrawn in the vast majority of patients,and treatment response was satisfactory.Systemic chemotherapy was given in 6 cases,and three of them died.Local radiation therapy was given in 4 cases,the tumor was significantly controled,and patients survived.Secondary liver transplantation was performed on 3 cases: 1 case died of recurrent lymphoma,and one case received partial hepatectomy and no lymphoma recurred.Conclusion For cases with obstructive symptoms of fever and chills associated with jaundice without reasonable explanation,LL-PTLD is suspected and diagnosed by liver biopsy.Basic treatments such as adjustment of immunosuppressive agents and anti-viral therapy are recommended as early as possible.Local radiation therapy is a treatment method of LL-PTLD,which can obtain a satisfactory therapeutic effect.

Objective To indentify the relationship between hepatitis B virus covalently closed circular DNA (HBV cccDNA) and postoperative recurrence of HBV in HBcAb positive liver donors by detecting HBV cccDNA in liver from HBcAb positive liver donors.Method Eighty-five of 1200 patients underwent liver transplantation for hepatitis B-related end-stage liver disease in our hospital from January 2007 to January 2010 were retrospectively analyzed.According to the situation of HBV infection in donor liver,the recipients were divided into 3 groups:(1) the experimental group (livers positive for HBcAb,and negative for HBsAg,n =40),(2) control group (livers positive for HBsAg,n =15),and (3) normal group (donor livers without HBV infection,n =30).HBV cccDNA of donors was detected by fluorescence quantitative PCR.Serum HBV and HBV DNA were regularly tested,and biopsy was done in those positive for HBsAg or HBV DNA to confirm HBV recurrence.The relationship between HBV cccDNA and postoperative recurrence of HBV in HBcAb positive liver donors was analyzed.Result The positive rate of cccDNA was 30％ in experimental group (12、40),73.3％ in control group (11/50),and 0 (normal group).The rate of HBV recurrence in experimental,control and normal groups was 10％ (4/40),80％ (12/15) and 3.3％ (1/30) respectively.The rate of HBV recurrence in the experimental group of cccDNA (+) and cccDNA (-) was 7.5％ (3/40),and 2.5％ (1/40).Conclusion The cccDNA in HBcAb positive donors may be one of the risk factors in HBV recurrence of hepatitis B-related liver transplantation patients.The screening of HBV cccDNA in the donor livers positive for HBcAb before liver transplantation is recommended to reduce the positive HBV recurrence and expand the pool of liver donors for patients with HBV-related liver disease.

BACKGROUND:Activation of Notch signaling plays a critical role in stem cel differentiation, and this effect seems to be cel-type dependent. Little is reported on the role of activation of Notch1 signaling in the differentiation of c-Kit+ bone marrow mesenchymal stem cels. OBJECTIVE:To analyze the influence of activation of Notch1 signaling on the differentiation of c-Kit+ bone marrow mesenchymal stem cels. METHODS:The Notch1 intracelular domain (N1-ICD) was obtained from the cDNA library by PCR and cloned intoBamHI/AgeI digested adenoviral GV314 plasmid to construct N1-ICD overexpressing shuttle plasmid, and the positive clones were verified by sequencing. N1-ICD shuttle plasmid and helper plasmids pBHGloxΔE1,3 Cre were used to co-transfect HEK293T cels to obtain N1-ICD overexpressing adenoviral particles (N1-ICD-Ad). The c-Kit+ subpopulation were isolated from bone marrow mesenchymal stem cels of the Sprague-Dawley rat femurviamagnetic activated cel sorting. After transfection of the c-Kit+ BMSCs with N1-ICD-Ad adenovirus, we assessed the activation of Notch1 signaling and differentiation in c-Kit+ bone marrow mesenchymal stem cels by quantitative RT-PCR and immunofluorescent staining. RESULTS AND CONCLUSION:N1-ICD coding sequence was successfuly generated from the cDNA library, and then was cloned into the linearized adenoviral vectors GV314. The resistant clones were verified by sequencing. With the assistance of packaging plasmids, recombinant N1-ICD-Ad adenovirus plasmids were successful packaged in HEK293T cels, and its title was 2×1012 PFU/L. c-Kit+ bone marrow mesenchymal stem cels with the purity of 91.6% were successfuly isolated from the bone marrow mesenchymal stem cels of the Sprague-Dawley rat femur. Compared with the blank and negative controls, N1-ICD-Ad infection in the c-Kit+ bone marrow mesenchymal stem cels led to substantial accumulation of N1-ICD in the cytoplasm and nuclei, significantly unregulated expressions of Hes1 (a downstream gene of Notch) and cardiomyocyte differentiation genes Nkx2.5 and cTnT, significantly increased the expression of von Wilebrand factor, an endothelial cel differentiation gene, and mildly increased the expression of smooth muscle22α, a smooth muscle cel differentiation gene. These experimental results indicate that the activation of Notch1 signaling contributes to multi-lineages differentiation of c-Kit+ bone marrow mesenchymal stem cels, and the construction of N1-ICD overexpressing adenoviral vector makes the foundation for further research on the role of Notch1 signaling in stem cel biology.

Objective To establish a high-performance induction culture system for Raw264.7 cells to differentiate into osteoclasts(OC)invitro by improving the cell culture program.Methods The Raw264.7 cells were cultured withα-MEM medium containing 50 μg · L-1 M-CSF, 100 μg · L-1 RANKL, and 1 × 10-8 mol · L-1 1α,25-(OH)2 D3 in 5% CO2 for 12 d at 37℃. The cells were digested transiently every time before the medium was changed after every three days. The morphologic changes of the Raw264.7 cells were observed by inverted microscope.The maturation and phagotrophic function of OC were identified by HE,TRAP,FITC-phalloidin staining and immunofluorescence.Results The cells remained to grow in single layers all the time in most areas of the well during the whole induction by the improved culture program. The observation results of inverted microscope and HE staining showed that the growth area of the polykaryotic OC reached to 70% of the well on day 1 2. FITC-phalloidin staining showed that in the maturation of the OC, the cluster-shaped podosomes in the pseudopodia gradually transformed into rings,which finally fused to form a large belt surrounding the periphery of the cytoplasm. The calcitionin receptor (CTR) expressed by OC was markedly enhanced compared with the precursor cells by immunofluroescence staining,and a large number of red granules appeared in the cytoplasm of OC with TRAP staining on day 1 2. These results comfirmed that the obtained OC were maturated and owned phagotrophic function. Conclusion A high-performance induction culture system for Raw264. 7 cells to differentiate into OC in vitro induced by combination of 50μg · L-1 M-CSF, 100 μg · L-1 RANKL,and 1 × 10-8 mol·L-1 1α,25-(OH)2 D3 is established by improving the cell culture program.

Objective·To analyze the choice of smoking cessation methods and its influencing factors in moderate-to-severe nicotine-dependent population.Methods·A retrospective clinical analysis was conducted for170 moderate-to-severe nicotine-dependent smoking-quitters (Fagerstr(o)m scale ≥ 4).Different smoking cessation methods (pharmacotherapy and non-pharmacotherapy) and abstinence rates of the selected population (7 d point prevalence abstinence rates and sustained abstinence rates 1,3,and 6 months after treatment) were compared and possible influencing factors were analyzed.Results·The differences in choosing the smoking cessation method in moderate-to-severe nicotine-dependent population with different ages,levels of education,and employment status were statistically significant (P=0.000).The Logistics regression analysis showed that the level of education was the major factor for choosing the smoking cessation method (P=0.006).The 7 d prevalence abstinence rate was much lower for pharmacotherapy than for non-pharmacotherapy 1 month after treatment (P=0.000) and was much higher for pharmacotherapy than for non-pharmacotherapy 3 and 6 months after treatment (P=0.000).The sustained abstinence rate was much higher for pharmacotherapy than for non-pharmacotherapy 3 and 6 months after treatment (P=0.002,P=0.000).Conclusion·For moderate-to-severe nicotine-dependent smoking-quitters,attention should be paid to the formulation and implementation of personalized strategies.The choice of smoking cessation methods depends on the age,level of education,and employment status.Reasonable pharmacotherapies can significantly increase the abstinence rate.

Objective To investigate the effect of amiloride on the invasion capacity of esophageal carcinoma EC9706 cell line in vitro and to elucidate its possible mechanism.Methods The invasion capacities of EC9706 cells pretreated with amiloride were measured by transwell chamber assay. The urokinase-type plasminogen activator (uPA) transcription were determined by RT-PCR.The protein expression of uPA were assessed by Western blot.Results After the EC9706 cells were pretreated with amiloride at different concentrations,the number of invaded cells was obviously less than those of control group with obvious dosage dependent pattern (96±7,78±6,57±6,33±4,15±3,F =43.46,P ＜ 0.01).The transcription levels of uPA mRNA and the protein expression levels of uPA in EC9706 cells decreased significantly compared with the control (mRNA:0.623±0.065,0.526±0.054,0.389±0.041,0.312±0.038,0.247±0.025,F =6.71,P ＜0.01; protein:0.732±0.064,0.644±0.057,0.533±0.058,0.391±0.036,0.267±0.043,F =6.71,P ＜0.01).Conclusion Amiloride inhibits the invasion capacity of esophageal carcinoma EC9706 cells.The mechanism might be associated with down-regulation of the expression of uPA.

Objective To investigate the expression and influence to tumor angiogenesis of urokinase-type plasminogen activator (uPA) and vascular endothelial growth factor (VEGF) in esophageal carcinoma. Methods The expression of uPA and VEGF in the tissue of normal (18 cases) and esophageal carcinoma (68 cases) were evaluated by SP immunohistochemistry, CD34 was detected as marking tumor microvessel density (MVD). uPA and VEGF expression were assessed as to the pathologically biological features of esophageal cancer and to the influence to tumor angiogenesis. Results The positive rates of uPA were 27.8 % (5/18) and 70.6 % (48/68) in the tissue of normal and esophageal carcinoma, respectively, there was significant difference in two tissues (x2 =11.63, P <0.05). The positive rates of VEGF were 22.2 % (4/18)and 63.2 % (43/68) in the tissue of normal and esophageal carcinoma, respectively, there was significant difference in two eissues (x2 =9.78, P <0.05). The expressions of uPA and VEGF in esophageal carcinoma were uniformity (x2 =9.72, P <0.05). The mean of MVD was 42.38±11.62. The positive rates of uPA and VEGF were higher in the high MVD group than those in the low MVD group (x2 =6.13, P <0.05, x2 =10.12, P <0.05,respectively). uPA and VEGF expressions in malignant tumors weren' t associated with age, gender and pathological types (P >0.05), but associated with clinical stage, histologic grading and lymph node metastasis (P <0.05). Conclusion Rising expression levels of uPA and VEGF are common in esophageal carcinoma. Altered expression of uPA and VEGF may contribute to tumor angiogenesis of esophageal carcinoma, whose overexpression indicate worse prognosis.

Objective:To compare the effects of Feishu(BL13)combined with Tianshu(ST25)and Feishu(BL13)alone on pulmonary function and tissue inflammation in asthma model rats. Methods:Forty-eight Sprague-Dawley rats were randomly divided into a normal group,a model group,a lung treatment group,and a lung-intestine treatment group according to the random number table method,with 12 rats in each group.Except for the normal group,rats in the other three groups were sensitized by ovalbumin followed by atomization stimulation to establish the asthma model.After the model was made successfully,rats in the lung treatment group were treated with acupuncture at bilateral Feishu(BL13)for 30 min;rats in the lung-intestine treatment group were treated with acupuncture at bilateral Feishu(BL13)and Tianshu(ST25)for 15 min,respectively,for 30 min in total.Acupuncture was performed once a day for 14 d.Rats in the other two groups did not receive any intervention.After intervention,the pulmonary function of each group of rats was measured by a pulmonary function tester.The histopathological changes stained by hematoxylin-eosin staining and the collagen deposition degree stained by Masson in lung tissue,as well as the inflammatory cells in the bronchoalveolar lavage fluid(BALF)stained by Wright's-Giemsa staining were observed under a light microscope;the levels of interleukin(IL)-4,IL-5,IL-13,IL-17,IL-25,IL-33,leukotrienes(LT),thymic stromal lymphopoietin(TSLP),and prostaglandin D2(PGD2)were detected by enzyme-linked immunosorbent assay. Results:Compared with the normal group,the peak expiratory flow(PEF),the dynamic lung compliance(Cdyn),the forced expiratory flow 25%(FEF25%),the forced expiratory volume in the first second/forced vital capacity(FEV1/FVC),and the maximum mid-expiratory flow(MMEF)in the model group were significantly decreased(P<0.01 or P<0.05);the lung resistance(RL),the collagen deposition,the IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 levels,and the neutrophil proportion in the BALF were increased significantly(P<0.01 or P<0.05).Compared with the model group,the FEF25%and FEV1/FVC in the pulmonary function were significantly increased(P<0.01,P<0.05),while the collagen fiber deposition and the levels of IL-4,IL-5,IL-13,IL-17,LT,TSLP,and PGD2 were significantly decreased(P<0.01 or P<0.05)in the lung treatment group;the PEF,FEF25%,and FEV1/FVC in pulmonary function were significantly increased(P<0.01 or P<0.05),while the RL and the collagen fiber deposition,as well as the neutrophil proportion and the levels of IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 in the BALF were significantly decreased(P<0.01 or P<0.05)in the lung-intestine treatment group.Compared with the lung treatment group,the IL-5 level in the rat lung tissue of the lung-intestine treatment group was decreased(P<0.05). Conclusion:Acupuncture in the lung treatment and the lung-intestine treatment groups improved pulmonary function,lung inflammation,and inflammatory cells in the BALF in asthma model rats;the lung-intestine treatment was better than lung treatment in improving the IL-5 level.The combination of Feishu(BL13)and Tianshu(ST25)was superior to Feishu(BL13)alone in the treatment of asthma in trend.

Objective: To determine the association between physical health fitness with Chinese reading ability of schoolaged children, so as to provide evidence for improving children s reading ability. Methods: A questionnaire survey was conducted among 1 923 school aged children in grades 2-6 in a primary school in Wuhan, Hubei Province, China. The questionnaire included basic demographic information and Dyslexia Checklist for Chinese Children and the Pupil Rating Scale Revised Screening. At the same time, participants underwent physical fitness tests which included an assessment of height, weight, and lung capacity, as well as a 50 meter run, sit forward bend, one minute skipping rope task, sit ups, and a 50 × 8 round trip. Results: A total of 59 children were identified with dyslexia. Normal children achieved higher scores than children with dyslexia in the total physical health score, as well as the one minute skipping rope score, one minute sit up score, and sitting forward score ( P <0.05). Multiple linear regression analyses showed that the reading ability of girls was higher than that of boys ( β =-3.04, P <0.01), and the children who regularly participated in more intense physical activity and who had higher fitness scores had a higher reading ability ( β =-1.68, -0.08, P <0.01). Children s reading ability increased significantly with parental educational level( P <0.05). Conclusion Gender, parents education level, physical exercise intensity, and children s physical fitness were identified as influencing factors of school age children s reading ability. A positive correlation was found between children s physical health level and reading ability.

Objective:To evaluate the effectiveness and safety of early exercise safety management in patients undergoing mechanical ventilation in intensive care unit (ICU).Methods:A prospective historical control observation was conducted. Forty-five patients with severe respiratory failure undergoing mechanical ventilation admitted to the ICU of Affiliated Hospital of Jining Medical University from April to June in 2019 were enrolled in the observation group and implemented early exercise safety management, including establishing multidisciplinary safety management team with ICU doctors, ICU nurses, respiratory therapists, rehabilitation therapists, dietitians and psychological consultants to jointly develop early exercise plan; equipping with sports and safety protection equipment; assessing the early exercise risks, formulating early exercise prescriptions; formulating the nutritional and psychological prescriptions; carrying out the propaganda and education in the early exercise process of patients and communicating with patients timely; strengthening team training. Other 45 patients with severe respiratory failure admitted for mechanical ventilation from January to March in 2019 were enrolled in the control group, whose gender, age and artificial airway conditions were matched with those in the observation group. Routine mechanical ventilation nursing was performed, including condition monitoring, medication nursing, environmental management and routine examination assistance. The improvement of depression, anxiety, comfort, lung function, and quality of life before and after nursing intervention were observed, and the occurrence of complications was recorded.Results:There was no significant difference in gender, age or artificial airway conditions between the two groups. Before nursing intervention, there was no significant difference in depression, anxiety, comfort, lung function and quality of life between the two groups. After the intervention of different nursing programs, the above conditions of the two groups were significantly improved, and the improvement was more significant in the observation group, which showed that the hospital anxiety and depression scale (HAD) score and forced vital capacity (FVC) of the observation group were significantly lower than those of the control group [HAD score: 10.80±2.54 vs. 17.51±3.66, FVC (L): 1.81±0.42 vs. 2.23±0.39, both P < 0.01], while the general comfort questionnaire (GCQ) score, forced expiratory volume in 1 second (FEV1), FEV1/FVC ratio, FEV1 percentage of predicted (FEV1%) and each dimension score of 36-item short form health survey (SF-36) scale were significantly higher than those of the control group [GCQ score: 110.87±5.33 vs. 96.93±3.02, FEV1 (L): 1.99±0.37 vs. 1.71±0.15, FEV1/FVC ratio: 0.88±0.04 vs. 0.84±0.03, FEV1%: (88.98±8.57)% vs. (80.41±4.45)%, mental function score: 49.74±9.88 vs. 40.17±8.95, physical function score: 27.65±9.46 vs. 20.32±9.53, social relationship score: 62.14±6.33 vs. 50.28±8.76, general health score: 38.61±8.48 vs. 30.63±8.93, all P < 0.01]. The overall incidence of complications in the observation group was significantly lower than that in the control group (24.44% vs. 57.78%, P < 0.01). Conclusions:Early exercise safety management scheme for patients undergoing mechanical ventilation in ICU can improve clinical efficacy and reduce complications. At the same time, it can further standardize the behaviors of medical staff and ensure the early exercise.