[Objective] To establish a method to seperate rat mesenchymal stem cells(rMSCs) from rat marrow, culture rMSCs in vitro, and study its biological characteristics. [Methods] rMSCs were seperated from rat marrow with wall-sticking method and expanding cultured in vitro. To draw the growth curves of cells at 1th, 3th, 5th passage, detect the surface antigens with flow cytometry. To evaluate the effect of fetal calf serum with different concentrations to rMSCs. [Results] There were approximately (5～6)?105 cells after primary culture and over (2~3)?108 rMSCs at 5th passage. The appearance of rMSCs is like shape of spear and its growth curves are like shape of S. The rMSCs were positive for CD90 and negative for CD45. [Conclusions] Cells seperated and cultured by the method established in this experiment were rMSCs. They have the biological characteristics of rMSCs, and are good resources for study in tissue engineering.

To sum up the experiences and results of surgical treatment for recurrent rectal carcinoma after rectectomy, pelvic exenterations were carried out for 49 patients with the disease and the survival rate was estimated. No death occurred in our group. The resect rate was 91 8%, and the operative complication rate was 12 2%. The 3 year survival rate was 85 5%, and the 5 year survival rate was 60 6%. Pelvic exenteration can improve the survival rate for recurrent rectal carcinoma after rectectomy. Urinary reconstruction and repair of pelvic peritoneum are important for reducing operative complications.

Objective To evaluate the efficacy of double filtration plasmapheresis (DFPP) in the treatment of patients with refractory rheumatoid arthritis (RA). Methods Eighty-two patients were randomly aesigned,42 to the DFPP group and 40 to the no-DFPP group. All patients previously experienced an incomplete response to 2-3 dis-ease-modifying antirheumatic drugs (DMARDs) and 1-2 nonsteroidal anti-inflammatory drugs (NSAIDs) or predni-sene. All patients received sulphasalazine (SASP,0.75 g three times daily) plus methotrexate (MTX, 10 mg orally once weekly). DFPP was performed once a week for 2-3 sessions. A total of 121 plasmapheresis procedures were per-formed in 42 patients. Control patients did not receive sham DFPP. The efficacy measures recorded one day after the final treatment and latest month in follow up for 12～24 months included the American College of Rheumatology 20% ,50% ,and 70% improvement criteria (ACR20, ACR50, and ACR70), the Health Assessment Questionnaire estimate of disability (HAQ); and the disease activity index. Results Patients in the DFPP group had ACR 20, ACR 50 and ACR 70 improvements of 100% ,92.9% and 81.0%,as compared with the patients in no-DFPP group 17.5% ,0,and 0 (P<0.001). Significant change from baseline was observed in HAQ scores in the DFPP group but not in the no-DFPP group (P<0.001). The changes from baseline in the disease activity scores were significantlygreater than in the no-DFPP group (P<0.001). Conclusion DFPP therapy significantly alters the signs and symp-toms of refractory RA. There are significant increases in physical function and improvement in quality of life.

Objective To observe the histological effect of acellular dermal matrix(ADM)on insufficiency in guide keratinized tissue regeneration. Methods 6 cases of single anterior tooth implantation in the hospital in 2016 were included in the study. 3 cases were treated with immediate tooth extraction and implantation. Bone substitution materials were grafted in the space between the tooth extraction socket and the implants. The keratin-ized tissue dehiscence was covered by double layers of acellular dermal matrix membrane(Heal-All?,ZH-BIO, China),which was fixed to the adjacent soft tissue by suturing. Another 3 cases were routinely treated with delayed implantation of single anterior tooth. All the cases were subject to harvesting of the cover soft tissues of implants with a punch 4 months later. The new grown soft tissues were histologically observed. Results All cases were sur-vived. The new grown keratinized tissues were observed. Conclusion Acellular dermal matrix can guide the aug-mentation of keratinized tissues.

OBJECTIVETo assess the prognostic value of CD20(+) tumor-infiltrating lymphocytes (TILs) in early-stage breast cancer.

METHODSParaffin sections were collected from 130 cases of stage I-III breast cancer undergoing surgery between January, 2000 and December, 2002 in our hospital. Immunohistochemistry was used to analyze mesenchymal CD20(+) TILs infiltration in the tumor and evaluate its association with the density of CD4(+) and CD8(+) TILs. The association of CD20(+) TILs was evaluated with the histopathologic features, overall survival (OS), distant disease-free survival (DDFS), and disease-free survival (DFS) of the patients.

RESULTSAggregations of CD20(+) lymphocytes were observed in 37.69% (49/130) of the cases. CD3(+) T cells were found to aggregate around CD20(+) B cell aggregations to form lymphoid follicle-like structures. The aggregations of CD20(+) TILs were positively correlated with the densities of mesenchymal CD8+ and CD4(+) TILs. Overall, CD20(+) TIL aggregations were not significantly correlated with the outcomes of the patients, but multivariate COX regressions suggested that CD20(+) TIL aggregations were positively correlated with DDFS (HR=0.251, 95% CI=0.071-0.894, P=0.033) and OS (HR=0.325, 95% CI=0.103-1.028, P=0.056) in hormone receptor-negative patients but not in the positive patients. Further analysis suggested that post-operative adjuvant endocrine therapy significantly improved the OS of patients positive for hormone receptors without CD20(+) TIL aggregations (P=0.001).

CONCLUSIONThe long-term therapeutic effects of adjuvant endocrine therapy are correlated with CD20(+) TIL aggregations to affect prognostic value of CD20(+) TIL aggregations in early-stage breast cancer patients.

Antigens, CD20 ; metabolism ; B-Lymphocytes ; cytology ; Breast Neoplasms ; immunology ; pathology ; Disease-Free Survival ; Female ; Humans ; Lymphocytes, Tumor-Infiltrating ; cytology ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; Survival Rate

OBJECTIVETo explore the relationship between a proliferation-inducing ligand (APRIL) expression in primary tumor foci of breast cancer and the patients' prognosis.

METHODSParaffin sections of surgical specimens were retrospectively collected from 130 stage I-III breast cancer patients who received surgery between January 2000 and December 2002 in our hospital. Immunohistochemistry was used to assess APRIL expression intensity in the tumor cells and density of interstitial APRIL-positive cells, and their association was analyzed with the density of interstitial CD4⁺ and CD8⁺ cells and with the histopathologic features, overall survival (OS), and disease-free survival (DFS) of the patients.

RESULTSAPRIL positive staining was found in the cytoplasm of the tumor cells, interstitial cells, and the extracellular matrix. APRIL intensity in the tumor cells was positively correlated with the density of interstitial APRIL-positive cells (P=0.009) and Ki67 (P=0.003). The density of interstitial APRIL-positive cells was positively correlated with the density of interstitial CD4⁺ cells (P<0.001) and CD8⁺ cells (P<0.001). In hormone receptor negative patients (ER- and PR-), multivariate COX regression identified the density of interstitial APRIL-positive cells as a positive prognostic factor for DFS (HR=0.313, 95% CI=0.107-0.920, P=0.035). CONCLUSIONSl APRIL is widely expressed in the interstitial immune cells in breast cancer. APRIL staining intensity in the tumor cells is positively correlated with tumor proliferation, indicating that the immune cells might promote tumor proliferation by secreting APRIL. A greater density of interstitial APRIL-positive cells is associated with a good prognosis in hormone receptor-negative patients.

Breast Neoplasms ; diagnosis ; metabolism ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Cell Transformation, Neoplastic ; Cytoplasm ; Disease-Free Survival ; Female ; Humans ; Immunohistochemistry ; Prognosis ; Retrospective Studies ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; metabolism

Objective:To examine the distribution of syphilis antibody in pregnant women and newborns and to explore how to optimize the existing syphilis screening process by setting the diagnostic gray area.Methods:The results of syphilis testing obtained from 119 531 pregnant women and 21 275 newborns from 2015 to 2018 by automatic chemiluminescent immunoassay (CLIA) and the re-examination results determined by Treponema pallidum particle agglutination (TPPA) and the rapid plasma reagin test (RPR) were retrospective analyzed. Data analysis was performed by Chi-square, Fisher′s exact test and Chi-square test for trend. Results:The positive rates of Syphilis specific antibody (TPAb) in clinical specimens from pregnant women and newborns were 0.69% (825/119 531) and 1.24%(264/21 275). The total re-examination positive rates were 0.32% (380/119 531) and 0.90%(191/21 275), and the suspicious syphilis prevalence rates in these specimens were 0.13% (161/119 531) and 0.31%(67/21 275), respectively. The suspicious syphilis prevalence rates in specimens of pregnant women from 2015 to 2018 and newborns increased year by year (χ 2=9.860, P=0.002; χ 2=5.311, P=0.021). With the elevation of the optical density value of samples to cut-off ratio (S/CO) value, positive coincidence rate of TPPA and TPAb in pregnant women and newborns increased significantly (χ 2=614.833, P<0.001; P<0.001). When the S/CO value in newborns exceeded 7.00 or the S/CO value in pregnant women exceeded 15.00, the effectiveness of TPAb results is equivalent to TPPA. The prevalence of suspected syphilis in pregnant women and newborns also increased with the increase of S/CO value (χ 2=323.059, P<0.001; P<0.001). When the S/CO value in newborns bellowed 3.00 or the S/CO value in pregnant women bellowed 5.00, the prevalence rate of suspected syphilis was 0%, which could preliminarily exclude syphilis infection. Conclusions:The prevalence rates of suspected syphilis in pregnant women was increasing during the recent years. It is necessary to further strengthen syphilis screening and intervention treatment in early pregnancy to improve the rate of eugenics. Being a primary screening method for syphilis in pregnant women and newborns, CLIA has high false positive rate. According to the gray area established in this study, the syphilis screening process can be optimized to prevent missed detection, which may reduce the false positive rate and avoid clinical misdiagnosis.

Objective: To investigate the clinical features and diagnostic bases of childhood leukoencephalopathy with cerebral calcifications and cysts (LCC). Methods: The clinical data involving manifestations and laboratory examinations of 4 children with LCC admitted to Beijing Children's Hospital Affiliated to Capital Medical University from 2012 to 2017 were retrospectively summarized. Each patient had a follow-up visit ranging from 4 months to 5 years and 9 months after initial examination. Results: Patients consisted of 2 males and 2 females, whose age of onset was respectively 2 years and 9 months, 6 years and 2 months, 7 years and 10 months, and 5 years and 1 month. The main clinical symptoms of these cases included headache, dizziness, partial seizure and claudication, and two of these cases had insidious onset. Cerebral calcifications and cysts with leukoencephalopathy were detected by neuroimaging in all patients. In addition, multifocal microhemorrhages and calcifications were observed by magnetic susceptibility-weighted imaging (SWI) series in 3 patients. Brain biopsy performed on 1 case disclosed a neuronal reduction in the cerebral cortex, loosening of focal white matter, multifocal lymphocyte infiltration, fresh hemorrhages, and gliosis, as well as angiomatous changes of blood vessels with hyalinized thicken-wall, stenotic or occlusive lumina and calcification deposits. The compound heterozygous mutations of n.*10G>A and n.82A>G in SNORD118 were identified in 1 case by target-capture next-generation sequencing. Sanger sequencing verified that the variant n.*10G>A was a novel mutation and it was of paternal-origin, while the variant n.82A>G was of maternal-origin, which had already been reported to be pathogenic to LCC. Follow-up study had shown continued partial seizure in 1 case and remissive claudication in another, while the remaining 2 cases had a relatively favorable outcome without obvious neurological symptoms at present time. Conclusions The clinical manifestations of LCC are nonspecific, and the onset of the disease tends to be insidious. The triad neuroimaging findings of cerebral calcifications, cysts and leukoencephalopathy are essential to the diagnosis of the disease, and the signals of microhemorrhages revealed by SWI series provide another eloquent reference for the diagnosis. As biopsy is invasive and usually unavailable in the early stage, gene assessment, instead of pathological data, should be the gold standard in the diagnosis of LCC.

Objective : To evaluate the clinical outcomes of custom all-ceramic crown fabricated in zirconia based on CT data in maxillary anterior implant tooth. Methods: 15 patients with single anterior tooth missing were applied to this treatment. Crown remodeling according to the opposite tooth, fabrication of the all-ceramic crown was finished based on the data capture via CT scan. The observation period extended 2-7 years on average. Results: The survival and success rates were 100%. Soft tissue was stabile in follow up photos. All of the patients were satisfied with the restorations. Conclusion The custom all-ceramic crown based on CT data showed good interface friendship with both of the hard and soft tissues and the aesthetics result is predictable.

Objective : To research the feasibility and preliminary clinical effect of an implant-supported fixed bridge based on interactions with the posterior interocclusal space deficiency. Methods: Four patients with multiple implant-supported fixed-bridge restorations for interocclusal space deficiency in posterior teeth were included in this study. The 8 total implant sites had an average interocclusal space size of 3.3 mm. Two abutments with an undercut area were performed, the fixed bridge was placed by rotating it without a common path of insertion, and the abutment screw was then tightened. In the production process, the interaction retention concept and methods were fully communicated to the technician. The abutments and bridges on the implants were placed, and the clinical effect was observed. Results: The prosthesis was fixed well and presented appropriate functioning. At the 3-month and 18-month follow-up examination, the prosthesis and abutments were not loose, and the abutments did not release or break. No swelling or tenderness was observed in the margin of the implants. Conclusion The interaction retention is a good method of resolving the problem of interocclusal space deficiencies in the posterior teeth.