Background and purpose:Microsatellite instability (MSI) status is commonly applied to predict the prognosis and chemosensitivity in stage Ⅱ and stage Ⅲ colorectal cancer patients. However, researches of its function on metastasis colorectal cancer are limited. This study investigated its value on prognosis and chemosensitivity in metastatic colorectal cancer (CRC) patients.Methods:We retrospectively investigated tumor tissues from metastasis CRC patients who were treated with oxaliplatin and 5-FU-based therapy regimens (FOLFOX and XELOX). Immunostaining of proteins of the mismatch repair genehMLH1,hMSH2,hMSH6 andhPMS2 was performed. Prognostic value and chemosensitivity in patients with MSI status were also determined.Results:Clinical features from 113 patients were analyzed. No cor-relation of overall survival (OS) and chemosensitivity with MSI status was found. We further investigated 79 patients with synchronous metastasis and palliatively tumor resection. Median progression free survival (PFS) from 22 MSI patients was significant longer than that in 57 MSS patients (19.9 monthsvs 7 months,P=0.005). No significant difference was seen in OS comparison (P=0.07). MSI status was also an independent prognostic factors of PFS by Cox multivariate analysis (MSS/MSI,HR=2.079,P=0.043). Moreover, in this group, MSI patients had improved disease control rate (59.1%vs 31.6%, P=0.025) in chemosensitivity analysis than MSS patients.Conclusion:A better PFS in MSI patients with synchronous metastasis and palliative tumor resection was found after treated with oxaliplatin and 5-FU-based therapy and a better chemosensitivity in MSI patients was also found.

Professor Cai Wanru believes that in the stable phase of chronic obstructive pulmonary disease, "phlegm" and "stasis" are intertwined, with "phlegm" as the main component. Disease is manifested mostly in the lungs, spleen and kidney, and along with the duration of the disease, the patients are prone to "sputum", "stasis" and other pathological products, so that acute aggravating symptoms are often induced due to the transformation from "phlegm" to "stasis". The clinical manifestations are mainly the lack of righteousness, and various symptoms due to phlegm stagnation and blood stasis. Therefore, Professor Cai believes that the thought of "preventive treatment of disease" should be throughout the treatment of the disease. The thought is based on the principle of the relationships between the five internal organs. According to the characteristics of disease transmission, combined with the syndrome differentiation of physical constitution, the importance of strengthening the body resistance and nourishing the vigour was emphasized. In terms of the methods of treatment, methods such as nourishing the lungs and tonifying the spleen, warming the kidney and tonifying qi are often applied, so the prescription is flexible and the compatibility is rigorous. In terms of the control of symptoms and prevention of acute exacerbations, it has a significant effect. Therefore, the thought of "preventive treatment of disease" proposed by Professor Cai Wanru has a practical value in the clinical treatment of chronic obstructive pulmonary disease, which is worthy of promotion.

OBJECTIVETo explore the regulatory effect of the GAGA element-related protein (GRP) on the Drosophila GAGA-dependent promoter activity in Jurkat cells.

METHODSDrosophila GAGA (dGAGA) factor (dGAF), GRP, and either the chloramphenicol acetyltransferase (CAT) reporter plasmid driven by the GAGA element-containing promoter of ftz gene or its mutant GAGA control were selectively co-transfected into Jurkat cells. Promoter activity analyses were performed by analyzing the RNA expression of the CAT in a real-time-RT PCR system, with pRC-CMV-betaGal co-transfected with the CAT reporter as a transfection efficiency control. Electrophoretic mobility shift assays (EMSA) were carried out to examine the binding profile of Jurkat nuclear extracts with a biotin labeled probe-containing dGAGA.

RESULTSIn Jurkat cells, GRP, either singly or combined with dGAF, elevated the activity of wild type dGAGA-containing ftz promoter dose-dependently in certain range. However, when the level of GRP was excessively high, it reduced or even fully inhibited the promoter activity of the ftz gene. On the contrary, either GRP or dGAF could not activate the ftz promoter with mutant GAGA. EMSA profile showed a specific band composed of the GAGA element and its binding proteins from Jurkat cells.

CONCLUSIONSHuman GAGA element-binding proteins exist in Jurkat cells. Its may either directly regulate the gene via GAGA elements or mediate the biphasic regulation of relevant gene in a GRP dose-dependent way.

Checkpoint Kinase 1 ; DNA-Binding Proteins ; genetics ; Drosophila Proteins ; genetics ; Gene Expression Regulation ; Humans ; Jurkat Cells ; Plasmids ; genetics ; Promoter Regions, Genetic ; Transcription Factors ; genetics ; Transfection

BACKGROUNDRapamycin (RAPA) is a relatively new immunosuppressant drug that functions as a serine/threonine kinase inhibitor to prevent rejection in organ transplantation. RAPA blocks activation of T-effector (Teff) cells by inhibiting the response to interleukin-2. Recently, RAPA was also shown to selectively expand the T-regulator (Treg) cell population. To date, no studies have examined the mechanism by which RAPA converts Teff cells to Treg cells.

METHODSPeripheral CD4(+)CD25(-) naive T cells were cultivated with RAPA and B cells as antigen-presenting cells (APCs) in vitro. CD4(+)CD25(-) T cells were harvested after 6 days and analyzed for expression of forkhead box protein 3 (Foxp3) using flow cytometry. CD4(+)CD25(+)CD127(-) subsets as the converted Tregs were isolated from the mixed lymphocyte reactions (MLR) with CD127 negative selection, followed by CD4 and CD25 positive selection using microbeads and magnetic separation column (MSC). Moreover, mRNA was extracted from converted Tregs and C57BL/6 naive CD4(+)CD25(+) T cells and Foxp3 levels were examined by quantitative real-time polymerase chain reaction (rt-PCR). A total of 1 x 10(5) carboxyfluorescein succinimidyl ester (CFSE)-labeled naive CD4(+)CD25(-) T cells/well from C57BL/6 mice were cocultured with DBA/2 or C3H maturation of dendritic cells (mDCs) (0.25 x 10(5)/well) in 96-well round-bottom plates for 6 days. Then 1 x 10(5) or 0.25 x 10(5) converted Treg cells were added to every well as regulatory cells. Cells were harvested after 6 days of culture and analyzed for proliferation of CFSE-labeled naive CD4(+)CD25(-) T cells using flow cytometry. Data were analyzed using CellQuest software.

RESULTSWe found that RAPA can convert peripheral CD4(+)CD25(-) naive T Cells to CD4(+)Foxp3(+) Treg cells using B cells as APCs, and this subtype of Treg can potently suppress Teff proliferation and maintain antigenic specificity.

CONCLUSIONOur findings provide evidence that RAPA induces Treg cell conversion from Teff cells and uncovers an additional mechanism for tolerance induction by RAPA.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antigen-Presenting Cells ; drug effects ; immunology ; metabolism ; B-Lymphocytes ; drug effects ; immunology ; metabolism ; CD4-Positive T-Lymphocytes ; drug effects ; immunology ; metabolism ; Cell Proliferation ; drug effects ; Dendritic Cells ; drug effects ; immunology ; metabolism ; Forkhead Transcription Factors ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-7 Receptor alpha Subunit ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Mitomycin ; pharmacology ; Polymerase Chain Reaction ; Sirolimus ; pharmacology ; T-Lymphocytes, Regulatory ; drug effects ; immunology ; metabolism

Objective: To analyze the epidemiological characteristics of coronavirus disease 2019 ( COVID-19 ) clusters in Lishui, so as to provide basis for the prevention and control of COVID-19 clusters. Methods: The data of COVID-19 clusters in Lishui from January 23 to March 29, 2020 were collected through China Disease Control and Prevention Information System-Public Health Emergency Information System, and analyzed time, space, scale, source of infection, exposure and transmission route by descriptive epidemiological method. Results: There were 31 cases in 8 clusters ( about 4 cases per cluster ), with no death. The report time was bimodal, peaked first from January 20 to February 10 with 4 clusters imported from domestic and peaked second from March 1 to 29 with 4 clusters imported from overseas. Qingtian County reported 4 clusters, Liandu District, Yunhe County, Qingyuan County and Jingning County each reported 1 cluster. Thirteen cases were restaurant employees, accounting for 41.94%. The cases were mainly occurred in the condition that exposed in the same family ( 6 clusters ), in the same dinner and car ( 1 clusters ), and in the same party ( 1 clusters ). The exposure modes that caused more cases infected were through the same family (9 cases) and through the same dinner and car ( 6 cases ). There were 3 clusters with first-generation cases, 3 clusters with second-generation cases and 2 clusters with third-generation cases. The recurrence rate of the 8 clusters ranged from 1.49% to 7.69%, with a median of 3.47%. Conclusions The COVID-19 clusters in Lishui imported from domestic in the early stage and later from overseas. Most cases were reported from Qingtian County, were engaged in catering business, and exposed by living with families.

Objective: To investigate the current status of prevention and control of occupational hazards in a city in 2017, to understand the capability to prevent and control occupational hazards and the level of occupational health supervision and management, and to propose measures to urge employers to assume the main responsibility for the prevention and control of occupational diseases. Methods: An analysis of the main factors influencing the prevention and control of occupational hazards in the city was performed to screen out six semi-quantitative assessment indicators (including the training of the main responsible persons and occupational health management personnel in companies) and four qualitative assessment indicators (including the coverage of supervision and inspection of occupational hazards performed by the district safety supervision department) , which could be used to measure the prevention and control effects of regional occupational hazards. Each indicator was scored. The typical investigation method was used to do data review and on-site inspection of 170 companies, 17 district-level occupational health supervision departments, and 16 sub-district (township) occupational health supervision departments in the city from October to December, 2017. The prevention and control of occupational diseases in each district was scored, and the completion rate and completion situation of each indicator in the city were analyzed. Results: The mean score of prevention and control of occupational hazards in the city was 84.9. The scores of two districts were relatively high, being 88.9 and 88.7, respectively; the scores of 9 (52.9%) districts were higher than 85. The pass rate of training for the main responsible persons and occupational health management personnel in companies in the city was 95.9%, the pass rate of occupational health training for workers was 84.7%, the pass rate of occupational health examination for workers was 96.5%, the pass rate for the inspection of occupational hazardous factors in workplaces was 95.3%, the pass rate for notifying occupational hazards in workplaces is 95.9%, and the pass rate for applying the warning signs of occupational hazards in workplaces was 76.5%. Conclusion The awareness of the importance of prevention and control of occupational hazards in all districts of the city has been increased, but the effectiveness of occupational health training for workers needs to be strengthened. The supply and demand of occupational health technical services are still not matched, and the ways and methods of occupational health supervision and inspection need to be improved.

Objective To construct a new conditionally replicative adenovirus (CRAds) targeting carcinoembryonic antigen (CEA) positive colorectal cancer cells. Methods The DNA fragment of the CEA gene promoter was amplified through PCR and cloned into the vector carrying fusion reporter gene EGFP-Luc to construct expression plasmid pCEA-EGFPLuc. The constructed plasmid pCEA-EGFPLuc was transfected into CEA positive and negative cells by liposome. The activity of CEA gene promoter was evaluated by detecting the expression of EGFP and luciferase activity. The conditionally replicative adenovirus Ad.CEA-E1A/CMV-TK carrying suicide gene HSVtk was constructed, in which the E1A gene was controlled under CEA promoter. CEA positive(Lovo and SW620)and negative tumor cells(HeLa) were infected with Ad.CEA-E1A/ CMV-TK. The selective cytotoxicity of Ad.CEA-E1A/CMV-TK and the synergistic effect of the virus with GCV in CEA positive tumor cells were evaluated by the expression of E1A, cytopathic effect and cell survival rate. Results CEA promoter possesses a good specificity as well as high activity. The expression of E1A only presented in CEA positive tumor cells. After infection with Ad. CEA-E1A/CMV-TK, the cell survival rates of Lovo and SW620 were (36.72±2.49)% and (39.82±4.76)%, respectively, significantly lower than that of Hela[(87.44±2.76)%1 (P<0.01). When combined with GCV, Ad.CEA-E1A/CMV-TK had better oncolytic effect on Lovo and SW620 cells, with cell survival rates of (17.26±3.65) % and (23.93±5.40) %, respectively, significantly lower than those without GCV[(36.72±2.49) % and (39.82±4.76) %, respectively] (P<0.01). Conclusion Ad. CEA-E1A/CMV-TK under the control of CEA promoter has selective cytotoxic effect on CEA positive colorectal cancer cells, and the effect can be enhanced when combined with GCV.

Objective:To investigate the feasibility and clinical value of MRI quantitative evaluation technique in detecting sternocleidomastoid muscle fibrosis in patients with nasopharyngeal carcinoma (NPC) after radiotherapy.Methods:From August 2019 to March 2021, 45 patients with clinically confirmed NPC after radiotherapy and 30 healthy controls who underwent physical examination in Lishui Hospital of Zhejiang University were enrolled in our study. According to the Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer (RTOG/EORTC) classification criteria of late radiation reactions respectively, the sternocleidomastoid muscle injury in the NPC group was divided into grade Ⅰ, Ⅱ and Ⅲ, which included 8, 32 and 5 patients respectively. All patients underwent T 1 mapping and T 2 mapping imaging of the neck. Firstly, the mapping images of sternocleidomastoid muscle between the two groups were analyzed and compared. Using NUMARIS/4 software of Siemens image post-processing workstation, the region of interest was manually drawn along the edge of sternocleidomastoid muscle at the level of laryngeal chamber in axial mapping diagram. Then, T 1 and T 2 values and the long and short diameters of sternocleidomastoid muscle were measured respectively. Finally, the differences of the parameters between the two groups were compared by independent sample t-test, Spearman rank correlation was used to analyze the relationship between the average T 1 and T 2 values of bilateral sternocleidomastoid muscles and the grade of late radiation injury. Results:Compared with the control group, the shape of sternocleidomastoid muscle in the NPC group was smaller in shape, with irregular edge and uneven increase of T 1 mapping color scale. There was no significant difference in muscle signal in T 2 mapping. The T 1 values of left and right sternocleidomastoid muscles in the NPC group were (1 524.7±97.6) and (1 496.5±93.2) ms respectively, which were significantly higher than those in the normal control group [(1 231.5±85.3) and (1 275.9±90.9) ms] ( P<0.05), and the T 2 values of left and right sternocleidomastoid muscles in the NPC group were (28.4±4.8) and (28.4±3.6) ms respectively, which were lower than those in the normal control group [(30.4±3.5) and (30.4±3.5) ms] ( P<0.05). The long and short diameters of bilateral sternocleidomastoid muscles in the NPC group were shorter than those in the control group ( P<0.05). The average T 1 and T 2 values of bilateral sternocleidomastoid muscles in NPC patients after radiotherapy were (1 510.6±95.4) and (28.4±4.2) ms respectively, The T 1 value was positively correlated with the classification of advanced radiation injury ( r=0.78, P<0.001), and T 2 value was negatively correlated with the level of advanced radiation injury ( r=-0.87, P<0.001). Conclusion:Mapping quantitative evaluation technique can noninvasively and objectively detect and evaluate sternocleidomastoid muscle fibrosis after NPC radiotherapy, which has potential clinical application value.

BACKGROUNDDNA hypomethylation of long interspersed nuclear elements-1 (LINEs-1) occurs during carcinogenesis, whereas information addressing LINE-1 methylation in Wilms tumor (WT) is limited. The main purpose of our study was to quantify LINE-1 methylation levels and evaluate their relationship with relative telomere length (TL) in WT.

METHODSWe investigated LINE-1 methylation and relative TL using bisulfite-polymerase chain reaction (PCR) pyrosequencing and quantitative PCR, respectively, in 20 WT tissues, 10 normal kidney tissues and a WT cell line. Significant changes were analyzed by t-tests.

RESULTSLINE-1 methylation levels were significantly lower (P < 0.05) and relative TLs were significantly shorter (P < 0.05) in WT compared with normal kidney. There was a significant positive relationship between LINE-1 methylation and relative TL in WT (r = 0.671, P = 0.001). LINE-1 Methylation levels were significantly associated with global DNA methylation (r = 0.332, P < 0.01). In addition, relative TL was shortened and LINE-1 methylation was decreased in a WT cell line treated with the hypomethylating agent 5-aza-2'-deoxycytidine compared with untreated WT cell line.

CONCLUSIONThese results suggest that LINE-1 hypomethylation is common and may be linked to telomere shortening in WT.

Cell Line, Tumor ; Child ; Child, Preschool ; DNA Methylation ; genetics ; Female ; Humans ; Long Interspersed Nucleotide Elements ; genetics ; Male ; Polymerase Chain Reaction ; Telomere ; genetics ; Wilms Tumor ; genetics

OBJECTIVETo study the hypoglycemic effect of the extract of B. polyandra (SHG).

METHODThe diabetic mice were induced by alloxan in ICR mice. The blood glucose concentration was measured by glucose oxidase method. The serum insulin level was determined by 125I-insulin radioimmunoassay kit. The hypoglycemic effect was evaluated by the levels of both fasting and no-fasting blood glucose. The effect on serum insulin level was estimated by the values of the blood insulin and the changes of the blood glucose induced by the glucose intraperitoneal injection. The effect on the glucose absorption was investigated by the oral sucrose or starch tolerance test.

RESULTBoth of the fasting and no-fasting blood glucose levels were decreased significantly by the treatment of 20 or 30 g raw materials crude drug x kg (-1) SHG orally for 7-10 d in ICR mice or in alloxan diabetic mice. In the oral sucrose tolerance test or oral starch tolerance test, the administration of SHG reduced significantly the peak value of the blood glucose and the area under the blood glucose-time curve (AUC) in normal or alloxan diabetic mice, respectively. These effects of SHG were similar to those of acarbose, a kind of alpha-glucosidase inhibitors. In the oral glucose tolerance test in normal and alloxan diabetic mice, SHG decreased both the blood glucose peak and the AUC induced by the glucose loading. But in the intraperitoneal injection glucose tolerance test the levels of insulin in both SHG and control mice were similar, however, the changes of the blood glucose level after the glucose-loading for 30 min in SHG mice was much lower than that in control mice.

CONCLUSIONWith the treatment of SHG, the fasting and no-fasting blood glucose concentrations were decreased and the glucose tolerance improved significantly in both normal and alloxan diabetic mice, and the inhibition of a-glucosidase might be one of its major mechanisms.

Administration, Oral ; Alloxan ; Animals ; Area Under Curve ; Blood Glucose ; analysis ; Diabetes Mellitus, Experimental ; blood ; chemically induced ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Glucose ; administration & dosage ; pharmacokinetics ; Glucose Tolerance Test ; Hypoglycemic Agents ; administration & dosage ; isolation & purification ; pharmacology ; Injections, Intraperitoneal ; Insulin ; blood ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; alpha-Glucosidases ; metabolism