OBJECTIVETo study the immuno-tolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells (BMSCs) in the allogeneic transplantation.

METHODSForty female C57BL/6 mice and forty male BALB/C mice were respectively used as donors and recipients in skin allogenic graft model. Forty male BALB/C mice were divided randomly into 4 groups: blank control group, CP group, BMSCs group , CP + BMSCs group, with 10 mice in each group. Before skin graft, high-dose abdominal injection of cyclophosphamide (200 mg/kg, 2 d, q. d.) was performed in recipient mice in CP and CP + BMSCs groups. On the transplantation day, a bonus of 2 x 10(6) BMSCs from the SD rat (SD-BMSCs) were injected through the tail vein in the BMSCs and CP + BMSCs groups. The observation and HE staining of skin grafts were used. The expressions of CD29, CD34, CD45 and CD90 of cells were analyzed by using flow cytometry in order to identify BMSCs. The CD4+, CD25+, Foxp3 and Treg cells of spleen were detected by flow cytometry. Cytokine in peripheral blood of recipient mice were measured by ELISA, including TGF-beta, IL-10 and IFN-gamma. T cells were co-cultured with 60Co-irradiated bone marrow MSCs from different individuals. The proliferative activity of T cells were evaluated with MTT assay.

RESULTSThe skin graft survival time was significantly prolonged in the CP + BMSCs group, as compared with that in the blank control group, the CP group, the BMSCs group, respectively. Cells cultured by whole bone marrow adherent cultivation showed CD29 (99.7%), CD44+ (96.7%), CD34- (1.6%), CD45- (1.3%). Compared with the control group and CP group, the ratio of the CD4+, CD25+, Foxp3+ and Treg cells significantly increased in the SD-BMSCs group and CP + BMSCs group (P < 0.05). Analysis of peripheral blood by ELISA showed significant high level of TGF-beta, IL-10 and low level of IFN-gamma in BMSCs group and CP group,compared with that in control group. When co-cultured with BMSCs from different individuals, T- lymphocytes proliferation decreased apparently in SD-BMSCs group and C57-BMSCs group (P < 0.05), but there was no significant difference between SD-BMSCs group and C57-BMSCs group (P > 0.05).

CONCLUSIONSThe immunotolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells in the allogeneic transplantation might be associated with its effect on the proliferation of Treg cells and increasing expression of TGF-beta and IL-10, decreasing expression of IFN-gamma.

Animals ; Bone Marrow Cells ; immunology ; Female ; Immune Tolerance ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; T-Lymphocytes, Regulatory ; immunology ; Transforming Growth Factor beta ; immunology ; Transplantation, Homologous

This study was purposed to set up real-time quantitative RT-PCR technique and to measure leukemia fusion gene transcripts in patients with chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL) and acute promyelocytic leukemia (APL). All plasmids containing the target gene sequences were constructed to establish the standard curves. A TaqMan based real-time quantitative RT-PCR was performed to measure aberrant fusion gene transcripts in 130 samples of peripheral blood (PB) or bone marrow (BM) from 49 patients with leukemia. The results showd that the BCR-ABL(P210) transcripts were detected in 28 (82.4%) out of 34 CML patients (the ratios of BCR-ABL(P210)/ABL varied from 0.01 to 3.19) and also in 2 (33.3%) out of 6 ALL patients. The BCR-ABL(P190) transcripts were detected in 2 (33.3%) out of 6 ALL patients. The BCR-ABL(P210) and BCR-ABL(P190) transcripts were both detected in 1 (2.9%) CML patients. The PML/RARalpha transcripts were detected in 7 (77.8%) out of 9 APL patients (the ratio of PML-RARa/ABL varied from 0.0014 to 3.199). The relative frequency of both bcr1 and bcr3 was 42.9%, while that of bcr2 was 14.3%. The transcript level of aberrant fusion gene varied from the clinical situation of patient. It is concluded that real-time quantitative PCR is a reliable, innovative and promising technology with high sensitivity and specialty. It has potential clinical value for defining diagnosis, typing tumor, selecting treatment, measuring the tumor load, monitoring fusion gene expression level and evaluating therapeutic strategies, which is worthy to be popularized.
Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; Leukemia ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo investigate the role of microtubule-actin crosslinking factor 1 (MACF1) in the response of glioma cells to temozolomide (TMZ).

METHODSTMZ was applied to a human gliomablastoma cell line (U87) and changes in the protein expression and cellular localization were determined with Western blot, RT-PCR, and immunofluorescence. The responses of the cells with MACF1 expression knockdown by RNA interference to TMZ were assessed. TMZ-induced effects on MACF1 expression were also assessed by immunohistochemistry in a nude mouse model bearing human glioblastoma xenografts.

RESULTSTMZ resulted in significantly increased MACF1 expression (by about 2 folds) and changes in its localization in the gliomablastoma cells both in vitro and in vivo (P<0.01). Knockdown of MACF1 reduced the proliferation (by 45%) of human glioma cell lines treated with TMZ (P<0.01). TMZ-induced changes in MACF1 expression was accompanied by cytoskeletal rearrangement.

CONCLUSIONMACF1 may be a potential therapeutic target for glioblastoma.

OBJECTIVETo study the potential risk factors of human infecting with Streptococcus suis.

METHODS1: M matched case-control study was conducted. 29 human cases of Streptococcus suis infection in the early phase were included in the case group, Patients' family members, neighbors and peoples who had worked together with patients to handle deceased or sick pigs in the last week were recruited as matched controls. There were 147 controls in total. Both cases and controls received questionnaire investigation including the ways to contact sick/dead pigs. Conditional logistic regression was employed to analyze matching data.

RESULTSAccording to the results of multivariate analysis, slaughtering (OR = 11.978, 95% CI: 3.355-42.756), carcasses cutting and processing (OR = 3.008, 95% CI: 1.022-8.849) sick/dead pigs were associated with cases related to human Streptococcus suis infection. The attributable risk proportion were 91.65% and 66.76% respectively. The other types of exposures to sick/ dead pigs, including feeding, selling, burying and eating, were not associated with the human Streptococcus suis infection in our study population.

CONCLUSIONSlaughtering, carcasses cutting and processing sick/dead pigs were important risky behavior for humans to be infected by Streptococcus suis.

Adult ; Aged ; Case-Control Studies ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Occupational Exposure ; adverse effects ; statistics & numerical data ; Risk Factors ; Streptococcal Infections ; epidemiology ; etiology ; microbiology ; Streptococcus suis ; physiology

OBJECTIVETo describe the clinical and epidemiological features of dead cases with human Streptococcus suis infections, and to find the target population for preventing death and the related indicators.

METHODSEpidemiological investigation on human Streptococcus suis infections was implemented used unified questionnaires. Analysis on dead cases and survival cases (as contrast) was done.

RESULTSThe population with highest fatality rate was in 40-49 age group. 97.37% of dead cases had toxic shock syndrome. The mean interval from onset to admission was 0.76 days, and the mean course was 2.11 days. The progression among dead cases was faster than that among survival cases. Chief clinical manifestations of dead cases that are more frequent than survival cases are purpura (73.68%), diarrhea (50.0%), dyspnea (21.05%), conjunctival congestion (34.21%), etc. Renal impairment and liver involvement in dead cases were more significant than that in survival cases. No significant difference between mean incubation period, exposure rates of main risk factors in dead cases and in survival cases was found.

CONCLUSIONPreventing toxic shock syndrome might reduce the fatality rate. The target population for preventing death is aged > or = 40. Liver function and renal function testing might be indicators for monitoring the progression of human Streptococcus suis infections.

Adult ; Aged ; China ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; Streptococcal Infections ; blood ; microbiology ; mortality ; pathology ; Streptococcus suis ; physiology ; Young Adult

Objective To explore the feasibility, safety and effectiveness of endoscopic submucosal dissection ( ESD) in the treatment of the synchronous multiple early cancer or precancerous lesions in esophageal and stomach. Methods A retrospective study was conducted on the data of 5 patients with synchronous multiple early cancer or precancerous lesions in esophageal and stomach who were treated by ESD in Endoscopy Center of Zhongshan Hospital from January 2008 to December 2013. The characteristics of lesions, and results of therapy and follow-up were analyzed. Results All 5 patients were male with mean age of 67. 8±13. 1 years. The mean size of esophageal lesions was 2. 1±0. 9 cm with 1 lesion located in the upper esophagus and 4 in the middle. The mean size of gastric lesions was 2. 5±1. 5 cm with 2 lesions in the antrum, 2 in the gastric angle and 1 in cardia. Lesions in 4 cases were removed at the same time and 1 at different times. All lesions achieved complete resection. Postoperative pathological results showed that there were 2 cases of esophageal precancerous lesions with gastric precancerous lesions, 2 cases of esophageal precancerous lesions with early gastric cancer, and 1 case of early esophageal cancer with early gastric cancer. The 5 patients with 10 lesions all achieved curative resection. Postoperative esophageal stricture occurred in 1 case, which was improved after dilation. Median follow-up time was 72 months, when 3 patients survived and 2 patients died. However, the cause of death was not associated with the treatment. Conclusion ESD is a minimally invasive endoscopic method for treating synchronous multiple primary early cancers in esophagus and stomach.

Objective: To evaluate the safety and efficacy of dental floss traction-assisted endoscopic submucosal dissection (DFS-ESD) for rectal neuroendocrine neoplasm (NEN). Methods: A retrospective cohort study was performed. Clinical data of rectal NEN patients undergoing ESD at Endoscopy Center of Zhongshan Hospital, Fudan University from January 2016 to December 2017 were retrospectively analyzed. Inclusion criteria: 1) age of 18 to 80 years old; 2) maximal diameter of lesions <1.5 cm; 3) tumor locating in the submucosa without invasion into the muscularis propria; 4) no enlarged lymph nodes around bowel and in abdominal cavity; 5) ESD requested actively by patients. A total of 37 patients were enrolled, including 23 male and 14 female cases with mean age of (56.0±11.3) years. All the lesions were single tumor of stage T1, and the mean size was 0.8±0.2(0.5-1.2) cm. Postoperative pathology revealed all samples as neuroendocrine tumors (NET). Seventeen patients received DFS-ESD treatment (DFS-ESD group) and 20 patient received conventional ESD treatment (conventional ESD group). In DFS-ESD group, after the mucosa was partly incised along the marker dots, the endoscopy was extracted, and the dental floss was tied to one arm of the metallic clip. When the endoscope was reinserted, the hemoclip was attached onto the incised mucosa; another hemoclip was attached onto normal mucosa opposite to the lesion in the same way. The submucosa was clearly exposed with the traction of dental floss and the resection could proceed. The conventional ESD group received the traditional ESD operation procedure. The operation time, modified operation time (remaining time after excluding the assembly time of dental floss traction in DFS-ESD group), en bloc resection rate, R0 resection rate, morbidity of operative complication, recurrence and metastasis were compared between two groups. Results: The average tumor size was (0.8±0.2) cm in DFS-ESD group and (0.7±0.2) cm in conventional ESD group (t=0.425, P=0.673). According to postoperative pathological grading of rectal neuroendocrine neoplasm, 13 were G1 and 4 were G2 in DFS-ESD group, while 17 cases were G1 and 3 cases were G2 in conventional ESD group without significant difference (P=0.680). There were no significant differences in baseline data between in the two groups (all P>0.05). All the basal resection margins were negative, the en bloc resection rate was 100% and the R0 resection rate was 100%. Pathological results showed tumor tissue close to the burning margin in 5 cases of conventional ESD group and in 2 cases of DFS-ESD group (P=0.416). The operation time was (17.9±6.6) minutes in conventional ESD group and (14.7±3.3) minutes in DFS-ESD group (t=1.776, P=0.084). The modified operation time of DFS-ESD group was (11.9±2.8) minutes, which was significantly shorter than (17.9±6.6) minutes in conventional ESD group (t=3.425, P=0.002). The hospital stay was (2.3±0.6) days and (2.0±0.5) days in conventional ESD group and DFS-ESD group, respectively, without significant difference (t=1.436, P=0.160). No patient was transferred to surgery, and no delayed bleeding or perforation occurred in either group. There was no recurrence or primary tumor-related death, and all the patients recovered well during a follow-up period of 14(1-24) months. Conclusion Dental floss traction-assisted ESD for rectal neuroendocrine neoplasm can simplify operation and ensure negative basal margin.

Objective:To evaluate the safety, efficacy and long-term survival of endoscopic submucosal dissection (ESD) for colorectal precancerous lesions and early cancer in the elderly over 80 years old.Methods:Clinical data of colorectal precancerous lesions and early cancer treated with ESD from January 2007 to December 2014 at Endoscopy Center of Zhongshan Hospital, Fudan University were retrospectively analyzed. A total of 721 patients with 778 lesions were included in this study. These patients were stratified by age: the super-elderly group (≥80 years old, 55 patients, 7.6%) and the non-super-elderly group (<80 years old, 666 patients, 92.4%). The outcomes of ESD, complication incidences, pathological characteristics, and long-term survival were compared between the two groups.Results:Except that the incidence of chronic concomitant diseases in the super-elderly group was significantly higher than that in the non-super-elderly group [54.5% (30/55) VS 31.5% (210/666), P<0.001], other baseline characteristics were not significantly different ( P>0.05). There were no significant differences in the complete resection rate [93.1%(54/58) VS 95.3%(686/720)], the R0 resection rate [89.7% (52/58) VS 93.2% (671/720)], the curative resection rate [84.5% (49/58) VS 90.3% (650/720)], the complication incidence [5.5% (3/55) VS 2.7%(18/666)], or the median hospitalization (2.98 days VS 2.54 days) between the two groups (all P>0.05). The three-year overall survival rates of the super-elderly group and non-super-elderly group were 95.8% and 98.0%, respectively, and the five-year overall survival rates were 85.1% and 97.4%, respectively. Conclusion:Colorectal ESD is safe and effective for elderly patients (age ≥80 years old) despite a significantly higher incidence of chronic concomitant diseases than that in the non-super-elderly patients.

The historical evolution， fermentation technology and key links of Sojae Semen Praeparatum （SSP） were sorted out by consulting ancient books and modern literature， and the influencing factors and control methods of quality were analyzed and summarized in order to provide reference for the quality control of SSP. After analysis， it was found that in the fermentation process of SSP， fermentation strains， miscellaneous bacteria， temperature and humidity were all important factors affecting the quality of SSP. The condition control of "post fermentation" process has been paid more attention to in the past dynasties. In addition， the delicious SSP recognized in ancient times should be made from mold fermentation， and the breeding and application of fermented mold may be the key point to solve the quality problem of SSP. Therefore， based on the evaluation indexes of SSP in the past dynasties， it is of great significance to study and optimize the technological conditions such as strain， temperature and humidity in depth to improve the quality of SSP.

Objective To investigate whether Hainan papayas has protective effects in an Aβ40-induced primary neuron injury model and elucidate the underlying molecular mechanism. Methods Cultured primary neurons from the dorsal root ganglia (DRG) of Sprague–Dawley (SD) rats were treated with 20 μM Aβ40 peptide, 100 μg/L Hainan papaya water extract, peptide plus extract, or culture medium for 24 h. Cell viability was measured by MTT assay, and neuronal apoptosis was evaluated by DAPI staining. ERK signaling pathway-associated molecule activation and changes in Bax expression were analyzed by Western blotting and immunofluorescence. Results A cell viability rate of (44.11 ± 6.59)% in the Aβ40 group was rescued to (79.13 ± 6.64)% by adding different concentrations of the extract. DAPI showed pyknotic nuclei in 39.5% of Aβ40-treated cells; the fraction dropped to 17.4% in the 100 μg/L extract group. ERK phosphorylation was observed in the Aβ40 group but was ameliorated by pretreatment with 100 μg/L extract. Hainan papaya water extract also prevented Aβ40-induced phosphorylation of MEK, RSK1 and CREB associated with ERK signaling and downregulated Bax expression in the neurons. Conclusion The results suggest that Hainan papaya water extract has protective effects on neurons; the mechanism may be related to suppression of ERK signaling activation.