Objective:To investigate the changes in adaptive phenotypes of Yersinia pestis ( Yp) during successive passages in macrophages. Methods:A Yp strain of 201-MI was induced by 50 successive passages of Yp 201 strain in Raw264.7 cells. Phenotypic characteristics of 201 and 201-MI strains were compared by analyzing their survival rates in macrophages, growth curves, biofilm formation abilities, acid and hydrogen peroxide-stress tolerance, and virulence to mammal cells (Raw264.7 and HeLa cells) and mice. Results:Comparing with 201 strain, 201-MI strain showed various phenotypic changes, including higher survival rate in Raw264.7 cells, faster growth in iron-deficient medium, higher tolerance to acid and hydrogen peroxide, decreased biofilm formation ability, and less damages to Raw264.7 and HeLa cells. More-over, 201-MI strain showed decreased virulence to mice in both subcutaneous and intraperitoneal challenges. Preliminary comparative genomics analysis revealed some indel and nonsense mutations in 201-MI strain, which might account for its phenotype changes.Conclusions:After successive passages in macrophages, Yp showed some phenotypic changes, which might reflect its adaptive evolution under the pressure of macrophages. Detailed multi-omics analysis would be of great help to understand the underlying genetic mechanisms of these changes, and the related Yp-macrophage interaction processes as well.

Objective: To discuss the effects of acupuncture plus Tuina (Chinese therapeutic massage) in intervening the mental development, motor function, cerebral hemodynamics, and muscle tone in kids with spastic cerebral palsy (CP). Methods: A retrospective analysis was conducted on 87 kids with spastic CP. They were divided into a control group and an observation group based on their original treatment plan. Kids in the control group received symptomatic treatment, motor development therapy, and Tuina treatment. The observation group was additionally given acupuncture treatment. One month was taken as a treatment course. After six courses, the efficacy, change in muscle tone, mental development, gross motor function, fine motor function, and the systolic velocity (Vs), mean velocity (Vm), and pulsatility index (PI) of the anterior cerebral artery (ACA), middle cerebral artery (MCA), and posterior cerebral artery (PCA) were compared between the two groups. Results: After treatment, the muscle tone score dropped in both groups (P<0.05) and was significantly lower in the observation group than in the control group (P<0.05). The observation group achieved more significant efficacy than the control group (P<0.05). After the intervention, the mental development index (MDI), psychomotor development index (PDI), gross motor function measure (GMFM) score, and fine motor function measure (FMFM) score all increased in both groups (P<0.05) and were notably higher in the observation group than in the control group (P<0.05). After treatment, the Vs and Vm of the ACA, MCA, and PCA were markedly higher in the observation group than in the control group (P<0.05), and the observation group was significantly lower than the control group comparing the PI of the ACA, MCA, and PCA (P<0.05). Conclusion: Based on the symptomatic treatment and motor development therapy, Tuina and acupuncture treatment can effectively treat spastic CP children, manifesting as reducing muscle tone, boosting mental development, and improving motor function and cerebral hemodynamics.

Objective:To investigate the clinicopathological characteristics of the T cell lymphomas with CD20 expression, and to better understand this rare entity.Methods:Two-hundred cases of T-cell lymphoma diagnosed in the Department of Pathology of the Affiliated Hospital of Qingdao University from November 2016 to February 2020 were examined, and 5 cases of CD20-positive T-cell lymphomas were identified and included. Combined with clinical data and review of the literature, the clinicopathological characteristics of the disease were analyzed.Results:The five patients were all male, and had an average age of 56 years (range, 47 to 64 years). There were 2 cases of monomorphic epitheliotropic intestinal T-cell lymphoma, 2 cases of mycosis fungoides (1 case was plaque stage and the other was tumor stage) and 1 case of indolent T-cell lymphoproliferative disorder of the gastrointestinal tract. Immunohistochemistry showed that all 5 cases expressed multiple T cell markers (CD3/CD4/CD5/CD7/CD8) and only one of B cell markers (CD20). Three of the 5 cases were negative for CD20 at the first diagnosis, while CD20 was diffusely positive on the second biopsy from the recurrence or progression of the disease, without expression of CD79a or PAX5. Epstein-Barr encoding region (EBER) in situ hybridization was negative in all 5 cases. T-cell receptor gene analysis showed monoclonal rearrangement of β or/and δ chains;Ig rearrangements were all polyclonal. None of the five patients were treated with rituximab, and 4 patients survived with disease and 1 patient survived without disease at the end of follow-up. Among them, the patient with mycosis fungoides at the cancerous stage has progressed rapidly and had poor quality of life.Conclusions:CD20-positive T-cell lymphoma is extremely rare. Its prognosis is closely related to the type of T-cell lymphoma, clinical stage and initial therapeutic effect. However, the expression of CD20 indicates the recurrence or progression of the disease, and the prognosis is relatively poor. When CD3 expression is absent in T-cell lymphoma, it is easy to be misdiagnosed as B-cell lymphoma. The combination of multiple immunohistochemical antibodies and molecular detection can improve the accuracy of diagnosis.

Objective To evaluate the effect of case-PBL method and case-PBL method combined with SPSS software on the teaching of medical statistics for medical undergraduates. Methods Students from two classes major in experimental medicine from Nantong University Xingling College were selected as Experimental Class 1 ( using case-PBL teaching ) and Experimental Class 2 ( using case-PBL teaching combined with SPSS software). Each class attended lessons for five times. The teachers would send cases for each lesson one week in advance, and students would form into groups to access information, learn and discuss independently. Representatives from groups spoke in class, members would complement and discuss, while teachers offered guidance, review, analysis and summary. We shortened the discussion time in the class, and added instructions on the operation steps of SPSS software for case analysis and how to interpret the analysis result correctly in Experiment Class 2 . The Control Class received traditional teaching . Questionnaires, process assessment and final assessment were used to evaluate teaching effectiveness. SPSS 21.0 software was used to implement variance analysis and chi-square test for comparison between groups. Results The scores of the two experimental classes were higher than those of the control class in the five process assessments ( P<0 . 05 ) . Experimental Class 2 was the best in three section process assessments including those of t-test, the rank sum test and the straight-line correlation (P<0.05). However, there was no difference in the theoretical test scores among the three classes (P=0.670). 94％-100％of the students in the two experimental classes believed that the teaching effects of the case-PBL method and case-PBL method combined with SPSS software were better than the traditional teaching method (P<0.05) in such areas as stimulating interest in learning, improving learning efficiency, etc. Students in Experimental Class 2 were more prone to think these methods improved teaching effect. Conclusion The case-PBL teaching combined with the application of SPSS software will further help students to systematically grasp the statistical knowledge and cultivate statistical practice ability . However , neither methods will improve students' theoretical test scores.

In this study, eight-month-old ectomycorrhizae of Tuber borchii with Corylus avellana were synthesized to explore the influence of T. borchii colonization on the soil properties and the microbial communities associated with C. avellana during the early symbiotic stage. The results showed that the bacterial richness and diversity in the ectomycorrhizae were significantly higher than those in the control roots, whereas the fungal diversity was not changed in response to T. borchii colonization. Tuber was the dominant taxon (82.97%) in ectomycorrhizae. Some pathogenic fungi, including Ilyonectria and Podospora, and other competitive mycorrhizal fungi, such as Hymenochaete, had significantly lower abundance in the T. borchii inoculation treatment. It was found that the ectomycorrhizae of C. avellana contained some more abundant bacterial genera (e.g., Rhizobium, Pedomicrobium, Ilumatobacter, Streptomyces, and Geobacillus) and fungal genera (e.g., Trechispora and Humicola) than the control roots. The properties of rhizosphere soils were also changed by T. borchii colonization, like available nitrogen, available phosphorus and exchangeable magnesium, which indicated a feedback effect of mycorrhizal synthesis on soil properties. Overall, this work highlighted the interactions between the symbionts and the microbes present in the host, which shed light on our understanding of the ecological functions of T. borchii and facilitate its commercial cultivation.
Colon ; Corylus ; Fungi ; Magnesium ; Mycorrhizae ; Nitrogen ; Phosphorus ; Podospora ; Rhizobium ; Rhizosphere ; Soil ; Streptomyces

Objective: To investigate the clinicopathologic characteristics, diagnostic features and prognosis of primary renal neuroendocrine carcinoma (NEC). Methods: The clinicopathologic data of eight cases of renal NEC was collected from January 2008 to December 2017 from Affiliated Hospital of Qingdao University. Immunohistochemical staining was performed, and follow-up information was analyzed, and the relevant literature reviewed. Results: The patients′ mean age at diagnosis was 45 years (range, 27-66 years); five were women, and three were men. The tumors located on the left side in five patients, and on the right side in three. Five cases were detected incidentally, and three patients presented with loin pain. Microscopically, these cases included five well-differentiated NECs (three carcinoids, two atypical carcinoids), two small cell NECs, and one large cell NEC according to the World Health Organization classification of 2016. The tumors infiltrated the renal capsule in six cases. Necrosis was found in five cases. Vascular invasion with tumor emboli was seen in three cases. Lymph node metastasis was identified in one case. Immunohistochemically, the expression rates of neuroendocrine markers CD56, chromogranin A (CgA) and synaptophysin (Syn) were 6/8, 4/8, and 8/8 respectively. Some of the NECs were positive for epithelial markers CKpan (6/8, with three cases showing focal positivity) and CAM5.2 (4/8) of variable degrees. The Ki-67 proliferation index was≤3% in the carcinoids; ≥50% in the small cell carcinoma and large cell carcinoma; and 5% and 8% for the two cases of atypical carcinoid, respectively. All cases were negative for EMA, CK7, CA9, CD10, CD117, PAX2, PAX8, WT1, p63, S-100 and TTF1. Three patients (two with small cell carcinoma and one with large cell carcinoma) died of extensive metastases at 3 months, 4 months and 9 months after operation, while five patients were well, without recurrence or distant metastasis for follow-up period of one to nine years. Conclusions Primary renal NEC is rare. Carcinoid is the most common histological type. The pathomorphological features and neuroendocrine markers (CD56, CgA, Syn), epithelial markers (CKpan, CAM5.2) and nephrogenic markers (PAX2, PAX8) are important for the diagnosis. Renal carcinoid tumors are indolent and prone to early metastasis, but are associated with prolonged survival. The small cell renal cell carcinoma and large cell carcinoma are highly malignant renal tumors with poor prognosis and short survival.

Objective To investigate the clinical pathological factors related to recurrence and metastasis of colorectal cancer patients treated with resection ,expect to improve the treatment effects .Methods The clinicopathologic and follow-up data of 212 patients with stage Ⅰ - Ⅲ colorectal cancer patients treated with resection from January 2009 to December 2013 were retrospec-tively analyzed by univariate and multivariate Logistic regression methods .Results Univariate analysis showed that T staging sys-tems ,gross type ,intestinal obstruction or perforation ,lymphatic and vascular invasion were correlated to recurrence of colorectal cancer after operation(P< 0 .05) ;age ,tumor size ,lymph node involvement ,T staging systems ,histological differentiation ,intestinal obstruction or perforation ,lymphatic and vascular invasion were correlated to metastasis of colorectal cancer after operation(P <0 .05) .Multivariate Logistic regression analysis showed that age ,tumor size and histological differentiation were independent factors for metastasis rate of colorectal cancer after operation(P< 0 .05) .There were no significant differences in Logistic regression analy-sis for recurrence(P> 0 .05) .Conclusion Age ,tumor size and histological differentiation are independent factors for metastasis rate of colorectal cancer after operation .

Objective To analyze the features of bone minimal density and bone quantitative ultrasound in men with different osteoporotic risk graded by osteoporosis self-assessment tool for Asian (OSTA).Methods After exclude the secondary osteoporosis,724 subjects over 50 years old were involved.The parameters of hight,weight,quantitative ultrasound index (QUI),QUS-T score were examined.The bone density (BMD) were measured by dual-energy X-ray absorptiometry (DXA)in 120 elderly men.All subjects were grouped into low (osteoporotic) risk group,moderate risk group and high risk group by OSTA index.120 subjects measured BMD were grouped into normal bone mass group,osteopenia group and osteoporosis group by WHO standard.The differences and correlation analysis in BMD,QUST,and QUI between these groups were analysed.Results The percents of low risk people,moderate risk people and high risk people were 56.4％ (408 cases),28.2％ (204cases),15.5％ (112 cases),respectively.There were 30.0％ (36 cases) normal bone mass people,58.3％ (70 cases) osteopenia people and 11.7％ (14 cases) osteoporosis people in groups measured BMD.QUS-T score,QUI were gradually decreased in groups of low risk,moderate risk and high risk (-0.56±1.09,-0.88±-1.28,-1.21±1.40; 98.47±19.04,92.62±22.49,87.68±24.43; all P ＜0.05) and had statistical significant differences between low risk and moderate risk,high risk groups,while had no differences between moderate risk and high risk groups.The femoral neck BMD and total BMD were gradually decreased in all the three groups (0.89±0.12,0.85±0.10,0.77± 0.10; 1.0±0.15,0.93 ± 0.11,0.83±0.1; all P＜0.01).Osteoporosis in the three groups were 3.4％ (2 cases),13.0％ (6 cases),37.5％ (6 cases),respectively and osteoporosis percents in moderate risk group and high risk group were higher compared with low risk group (x2=11.77,P＜0.01).QUS-T score and QUI decreased gradually in groups of normal mass,osteopenia and osteoporosis (0.99±0.08,-0.70±1.07,-1.96±0.73; 109.26±17.05,96.15±18.20,72.54±10.00; F=10.47,11.73,all P＜ 0.01).Except for lumbar BMD,a positive linear correlation emerged between OSTA and QUS-T score,QUI,hip BMD(all P＜0.01).The values of R with femoral neck BMD,torch BMD and total hip BMD were 0.45,0.38,0.39,respectively.And the same value with QUS-T score and QUI was 0.23.Conclusions With the decreasing of OSTA index,risk of osteoporosis is increased and QUS-T score,QUI and BMD are decreased gradually.There are positive linear correlation between OSTA index and QUS-T score,QUI,hip BMD.

Objective To inspect the efficacy and mucosa healing condition of infliximab with azathioprine combination therapy in Crohn's disease (CD) and its correlation with prognosis.Methods A total of 20 active CD patients who received infliximab and azathioprine combination therapy at The First Affiliated Hospital of Sun Yat-sen University were objects of this study.The clinical efficacy was evaluated at 10 weeks,30 weeks,54 weeks and 2 years respectively according to CD activity index.The efficacy was evaluated under endoscopy at 10 weeks,30 weeks,54 weeks and 2 years respectively according to mucosal response situation under endoscopy.The data were analyzed by analysis of variance or Fisher's exact test between two groups.The factor affecred mucosal healing was analyzed by Logistic regression analysis.Results The clinical remission rate of patients without steroid at week 10,30,54 and 2 year were 12/20,16/20,15/20 and 15/20 respectively.Mucosal healing rate at week 10,30 and 54 weeks were 8/20,12/20 and 10/20 respectively.Logistic regression analysis indicated that age was the only factor affected mucosal healing at 30 weeks (OR =0.774,95％ CI:0.630 to 0.950).There was significant differences in clinical remission between mucosa response patients and invalid under endoscopy at 30 weeks and 2 years without steroid (at 30 weeks:14/14 vs 2/6; at 2 years:14/14 vs 1/6; all P＜0.01).Infliximab were withdrawn in 4 of 16 patients who was in non-steroid clinical remission at 30 weeks,and the other 12 patients were continued with infliximab therapy.There was no significant difference in non-steroid clinical remission rate (4/4 vs 11/12) and mucosa healing rate (2/4 vs 7/12) between withdrawal and continue of infliximab therapy (all P＞0.05).Conclusions Infliximab with azathioprine combination therapy can effectively promote and maintain mucosa healing in CD.The mucosa response patients can maintain long time non-steroid clinical remission.

ObjectiveTo explore the role of BTBD10 overexpression in the proliferation of insulinoma cell line INS-1and its mechanism. MethodsThe recombined expression plasmid of pcDNA4.0-BTBD10 was constructed by gene cloning technique and was transfected into INS-1 cell by lipofectamine 2000. The stable overexpression BTBD10 of INS-1cell was selected at 48th hour after transfection.INS-1 cell proliferation activity was measured by MTT method.The expression of BTBD10,protein kinase B(Akt),phospho-Akt(p-Akt),mammal target of rapamycin (mTOR) and phospho-mTOR (p-mTOR) were determined by Western blot.ResultsThe stable overexpression BTBD10 of INS-1 cell wassuccessfullyconstructed.OverproductionofBTBD10promotedbetacellproliferation.The phosphorylation of Akt and mTOR was increased and the ratio of p-Akt/Akt and p-mTOR/mTOR was enhanced in the INS-1 overexpressed by BTBD10.But the expression of total Akt and mTOR presented no obvious changes. Conclusion The overexpression BTBD10 of INS-1 cell could activate of Akt/mTOR signalling pathway via stimulating phospho-mTOR and Akt,and enhance overall cell protein translation,so as to promote proliferation of INS-1 cell.