[Summary] A total of 215 pregnant women at mid-gestation were enrolled, and their serum 25-hydroxyvitamin D levels were measured. Of them, 38 (17. 7%) were classified as cases with vitamin D sufficiency, 70 (32. 5%) as insufficiency, and 107 (49. 8%) as deficiency. Compared with the sufficient group, parathyroid hormone level increased and osteocalcin level decreased significantly in the deficient group. Therefore, vitamin D deficiency is highly prevalent among pregnant women and is associated with their potential disorder of bone turnover.

OBJECTIVEIn order to deeply understand the effects of nucleoside analogues to HBV DNA polymerase area, which initiate a state of drug resistance due to HBV genome mutation.

METHODSUsing PCR product direct sequencing method to test the gene sequence of P area of HBV genome of the patients who taking nucleoside analogues and showing signs of virologic breakthrough.

RESULTSMore resistance mutations can be find in HBV DNA polymerase area, which causing clinically cross drug resistance and multiple resistance.

CONCLUSIONWe should choose higher genetic barrier for the initial treatment. And the patients long-term using of nucleoside analogues, should be regularly monitored serum HBV DNA level, in order for early detection of virological breakthrough and timely intervention.

Adult ; Drug Resistance, Viral ; genetics ; Female ; Genome, Viral ; genetics ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Mutation ; genetics ; Polymerase Chain Reaction

Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.

BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) have low immunogenicity and it is unclear whether insulin producing cells (IPCs) that differentiate from hUC-MSCs have low immunogenicity. OBJECTIVE:To investigate the immunogenicity of IPCs differentiating from hUC-MSCs in vitro and after IPCs transplantation into the host. METHODS: (1) The hUC-MSCs were induced to differentiate into IPCs according to the modified scheme. Flow cytometry assay was used to detect the immunophenotype and apoptotic rate of IPCs in a cytotoxicity test. (2) Cell counting kit-8 was used to detect the proliferative capacity of human peripheral blood mononuclear cells in the one-way mixed lymphocyte assay. (3) The IPCs were then transplanted into the abdominal cavity and left renal capsule of mice, and then the infiltration of immune cells was detected by flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: The IPCs highly expressed HLA-ABC and lowly expressed HLA-DR, CD40 and CD80. The apoptosis rate of IPCs increased with the increase of pre-sensitized splenocytes in the cytotoxicity test. In the one-way mixed lymphocyte assay, IPCs inhibited the proliferation of human peripheral blood mononuclear cells when the target ratio was 10:1 and 50:1. After IPCs transplantation, the number of lymphocytes was increased in the transplanted site. In summary, our results show that IPCs that differentiate from hUC-MSCs maintain low immunogenicity in vitro,but have some immunogenicity after transplantation into the host due to microenvironment changes.

BACKGROUND: Mesenchymal stem cells are a kind of adult stem cells with self-renewal and multi-directional differentiation potential. These cells have the functions of immunoregulation, regulation of cell growth and repair of injury. In recent years, it has been found that when inflammatory injury occurs, mesenchymal stem cells can regulate the secretion of inflammatory factors, and function on the damaged region, thereby repairing and improving tissue damage caused by inflammation. OBJECTIVE: To review the immunoregulatory effects of mesenchymal stem cells on T lymphocytes, B lymphocytes, dendritic cells and natural killer cells when the body is in an inflammatory state. METHODS: A computer-based search of PubMed and CNKI was performed for articles published from 2000 to 2018 using the keywords of"inflammatory microenvironment, mesenchymal stem cell, immune response, T cell, B cell, DC, NK cell" in English and Chinese, respectively. Seventy-six articles related to the topic and with reliable arguments were finally included in result analysis. RESULTS AND CONCLUSION: The interaction between mesenchymal stem cells and inflammatory cells determines the result of tissue damage repair. In an inflammatory state, the biological characteristics of mesenchymal stem cells will undergo certain changes, but mesenchymal stem cells can still exert immunomodulatory effects by secretion of various soluble cytokines or via cell contact. There are still many problems to be further explored to facilitate better clinical application of mesenchymal stem cells.

Objective: To evaluate the choice of surgical treatment of ductal carcinoma in situ (DCIS) and its impact on long-term outcomes. Methods: A retrospective analysis of the clinicopathological features and treatment protocol of DCIS patients who underwent surgical treatment in Comprehensive Breast Health Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine from January 2009 to August 2016 was done. The factors which could affect surgical treatment were analyzed by χ² test and Logistic regression. Survival analysis were performed between different surgical approaches. Kaplan-Meier survival curves and Log-rank tests demonstrated the distribution of disease free survival and overall survival. Results: A total of 526 patients were enrolled in this study, 405 cases (77.0%) underwent mastectomy, 121 cases (23.0%) underwent breast-conserving surgery, of which 88 cases received radiotherapy after breast-conserving surgery. It was shown by univariate and multivariate analysis that age>50 years (OR=0.631, 95% CI: 0.413 to 0.965, P=0.034), first symptom of nipple discharge (OR=0.316, 95% CI: 0.120 to 0.834, P=0.020), excision biopsy (OR=1.831, 95% CI: 1.182 to 2.835, P=0.007) and tumor size >3 cm (OR=0.422, 95% CI: 0.206 to 0.864, P=0.018) were significantly correlated with choice of surgical treatment for breast lesions. Axillary lymph node dissection was performed for 118 cases (22.4%), with sentinel lymph node biopsy for 327 cases (62.2%), and none for 81 cases (15.4%). There was significant statistical difference in the choice of axillary lymph node management in patients of different age (χ²=8.124, P=0.017), biopsy type (χ²=35.567, P=0.000), breast operation type (χ²=149.118, P=0.000) and tumor size (χ²=13.394, P=0.010). The 5-year disease free survival rates was 95.7%, 89.6% and 100%, respectively, for mastectomy group, breast-conserving surgery group and breast-conserving surgery plus radiotherapy group. And the 5-year overall survival rates for three groups were 99.0%, 100% and 100%. The differences were not statistically significant (P=0.427, 0.777). Conclusions For DCIS patients, age, first symptom and tumor size are independent predictors of breast surgery. The choice of axillary lymph node surgery is influenced by age, biopsy, operation type, and tumor size. Different surgical treatment options has no significant effect on disease-free survival and overall survival in DCIS patients.

PURPOSE: The purpose of this study was to investigate the changes of tumor infiltrating lymphocytes (TILs) between core needle biopsy (CNB) and surgery removed sample (SRS) in early stage breast cancer patients and to identify the correlating factors and prognostic significance of TILs changes. MATERIALS AND METHODS: A retrospective study was carried out on 255 patients who received CNB and underwent surgical resection for invasive breast cancer. Stromal TILs levels of CNB and SRS were evaluated respectively. Tumors with ≥50% stromal TILs were defined as lymphocyte-predominant breast cancer (LPBC). Clinicopathological variables were analyzed to determine whether there were factors associated with TILs changes. Log-rank tests and Cox proportional hazards models were used to analyze the influences of TILs and TILs changes on survival. RESULTS: SRS-TILs (median, 10.0%) were significant higher than CNB-TILs (median, 5.0%; p<0.001). Younger age (<60 years, p=0.016) and long surgery time interval (STI, ≥4 days; p=0.003) were independent factors correlating with higher TILs changes. CNB-LPBC patients showed better breast cancer-free interval (BCFI, p=0.021) than CNB-non-LPBC (CNB-nLPBC) patients. Patients were categorized into four groups according to the LPBC change pattern from CNB to SRS: LPBC→LPBC, LPBC→nLPBC, nLPBC→LPBC, and nLPBC→nLPBC, with estimated 5-year BCFI 100%, 100%, 69.7%, and 86.0% (p=0.016). nLPBC→LPBC pattern was an independent prognostic factor of worse BCFI (hazard ratio, 2.19; 95% confidence interval, 1.06 to 4.53; p=0.035) compared with other patterns. CONCLUSION: TILs were significantly higher in SRS than in CNB. Higher TILs changes were associated with younger age and long STI. Changing from nLPBC to LPBC after CNB indicated a worse BCFI, which needs further validation.
Biopsy, Large-Core Needle ; Breast Neoplasms ; Breast ; Humans ; Lymphocytes, Tumor-Infiltrating ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Sexually Transmitted Diseases

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Receptor, ErbB-3 ; biosynthesis ; genetics

OBJECTIVETo evaluate the effects of a solitary megadose protocol of ionizing radiation (IR) to parotid gland on the structured and function changes of bilateral parotid glands in miniature pig.

METHODSFourteen minipigs were subjected to either 15 or 20 Gy to one parotid gland with a linear accelerator, while another four minipigs served as non-IR controls. Salivary flow rates and salivary chemistries were measured pre-IR, and 4 and 16 weeks post-IR. A quantitative assessment of gland weight and acinar area, and detailed serum chemistry and hematological analyses, were also performed.

RESULTSParotid gland weights were significantly decreased in the 15 and 20 Gy groups at 4 and 16 weeks post-IR. The acinar cell area in glands of both IR groups was significantly reduced. Parotid flow rates decreased by 60% with 15 Gy at 16 weeks post-IR. In the 20 Gy group, salivary flow rates were reduced by 80% at 16 weeks post-IR. Additionally, parotid flow rates significantly reduced in contralateral glands with 20 Gy at 16 weeks, while structure and weight did not changes in parotid glands.

CONCLUSIONStructural changes in salivary gland parenchyma occurred relatively early after IR, while the alterations in salivary output were relatively delayed. Further, reductions in salivary flow were not proportional to acinar cell area loss. There isn't a significant structured change of contralateral glands, but significant reduction of parotid flow rate at this time.

Animals ; Parotid Gland ; radiation effects ; Swine ; Swine, Miniature

Objective To evaluate the AChE and nAChR in the NMJ and the morphology of the muscle in the bilateral triceps surae after the unilateral shock wave therapy. Method 60 male New Zealand rabbits weighing (2± 0.2) Kg were used in this study. Two thousand shock waves at an energy flux density of 1.5bar and the frequency of 10Hz were applied to their left calf muscles. Divided into six groups, both sides of the triceps muscle of calf were taken out on the day of the shock wave and the1,2,4,6 and 8 weeks after the treatment. HE staining was used to observe the morphological changes of muscle tissue and the average optical density was measured after AChE stain so as to calculate the receptor count after Acetylcholine receptor immunohistochemistry. Result No abnormal morphological abnormalities were observed in all rabbits. In the first five groups, the AChE was significantly higher in the side of the shockwave treatment compairing with the control side (<0.05),slow decrease after 1 week after the treatment. In the first five groups, the nAChR was significantly lower in the side of the shock wave treatment compairing with the control side ( <0.05), and gradually increased to normal after 8 weeks. Conslusion Suitable dose of shock wave will not have a greater impact on morphology of muscle tissue. After the shock wave treatment, the amount and degree of stimulate of muscle cells were decreased, and the production of action potentials was reduced. While the experimental side AchE and AchR in shock wave treatment day to 8 weeks after treatment showed a significant trend to normal, it shows that the effect of shock wave on NMJ is transient and reversible.