Objective To investigate the effects of the expressions of thymidine kinase 1 (TK1) and Ki67 alone or their combination on the recurrence and metastasis of breast cancer.Methods Sixty-five samples were selected from the Second Affiliated Hospital of Guangzhou Medical University from March 2005 to June 2007,which were resected by surgical operation and confirmed as breast carcinoma by pathology.They were individed into two groups including 37 cases with recurrence or metastasis in 5 years (group A),28 cases without recurrence or metastasis in 5 years (group B).The expressions of TK1 and Ki67 in the two groups were detected by immunohistochemical staining assay.Then,Kaplan-Meier assay was used to describe survival curve.Results The positive expression rate of TK1 in group A was 91.8％,which was dramatically higher than that in group B 67.8％ (x2 =6.116,P =0.013).The positive expression rates of Ki67 in group A and B were 78.4％ and 42.9％ (x2 =8.635,P =0.003).The positive expression rates of TK1 combined with Ki67 in group A and B were 67.6％ and 39.3％ (x2 =5.159,P =0.023).Moreover,disease free survival of patients with positive expression of TK1 combined with Ki67 decreased significantly,compared with patients with positive expression of TK1 or Ki67 alone (x2 =6.137,P =0.046).Conclusion Positive expression of TK1 combined with Ki67 is the high risk factor of the reccurence or metastasis of breast carcinoma,and indicates poorer prognosis compared with positive expression alone.

Objective To construct humanized monoclonal antibodies against CD 20 and check their affinity to CD 20 antigen and their anti-tumor activity.Methods Based on the computer model , human IgG1 candidates closest to rituximab in crystal structure were selected in the Protein Data Bank ( PDB) .With the selected human IgG 1 candidates as the frame , we modified and transplanted the complementarity determining region ( CDR) of rituximab .First,the target gene fragments were obtained by overlapping PCR.Then, the sequences of the light chains(L) and the heavy chains(H) were inserted in-to the pcDNA3.3 and pOptiVEC vectors.Next, the constructed clones were transfected into 293F cells through transient transfection.After a large-scale cell culture, the mAb was purified by affinity chromatography rProtein A column.The puri-ty and expression level of the humanized antibodies was tested by sodium dodecyl sulfate ( SDS)-polyacrylamide gelelectro-phoresis(PAGE).The affinity of the humanized antibodies to CD20 was assessed with Fortebio assay.Finally, the anti-tumor activity of the constructed antibodies was detected by checking the tumor growth inhibition of the nude mice transplan-ted with tumor .Results Three humanized monoclonal antibodies against CD 20 were expressed and purified successfully . In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25 ×103 and 55 ×103 , respectively.The band size of the antibodies matched the expected value.Fortebio assay revealed that the humanized antibodies could bind to CD20 with high affinity (rituximab:6.48 ×10 -9mol/L, L4H7:1.91 ×10 -9mol/L, L5H5:7.35 ×10 -10mol/L,and L5H7:1.91 ×10 -9mol/L).The tumor growth inhibition experiment showed that the anti-tumor activity of L5H7 mAb was better than that of rituximab .Conclusion Three humanized monoclonal antibodies against CD 20 have been successfully construc-ted and expressed.L5H7 mAb possesses high affinity for CD20 and a good ability to kill tumor cells.

Objective:To understand the management status of breast cancer-related lymphedema, and to provide a basis for the formulation of prevention and treatment plans for breast cancer-related lymphedema.Methods:Using accidental sampling, a questionnaire designed by the research team was used to survey nurses in 58 hospitals in Jiangsu Province.Results:This study surveyed a total of 58 hospitals, of which 53 were general hospitals and 5 specialized hospitals. Only 11 hospitals (18.97%) had treatment rooms dedicated to conservative treatment of lymphedema, 9 hospitals (15.52%) had opened lymphedema clinics, and 15 hospitals (25.86%) had professional international lymphedema therapists. All the 58 hospitals had different degrees of prevention and management of breast cancer-related lymphedema.Conclusions:The comprehensive management of lymphedema has not yet been achieved, and the management standards of lymphedema also need to be further improved. It is important to establish a unified and standardized management plan and practice standards.

Objective: To investigate the occupational delay of gratification among community healthcare workers and its influencing factors, so as to provide insights into the sustainable development of primary healthcare personnel. Methods: The in-service community healthcare workers from 5-7 community health service centers in 9 cities (prefectures) of Guizhou Province were selected using a multi-stage stratified random sampling method. Gender, age, and educational level and other basic information were collected through questionnaire surveys. The status of occupational delay of gratification was investigated using the Occupational Delay of Gratification Scale. Multiple linear regression model was used to analyze the influencing factors of occupational delay of gratification. Results: A total of 2 076 respondents were surveyed, including 367 males (17.68%) and 1 709 females (82.32%). There were 112 respondents (5.39%) with secondary vocational school degree or below, 872 respondents (42.00%) with junior college degree, 1 087 respondents (52.36%) with bachelor's degree, and 5 respondents (0.24%) with master's degree or above. There were 665 respondents (32.03%) with managerial positions. The monthly income of 1 705 respondents (82.13%) was ≤5 000 Yuan. The total score of occupational delay of gratification was (33.22±4.33) points, and the total average score was (2.77±0.36) points. The average scores of work delay, career delay and persistence were (2.67±0.48), (2.96±0.45) and (2.75±0.46) points, respectively. Multiple linear regression analysis identified educational level (junior college, β=0.089; bachelor's degree, β=0.088), management position (not have, β=-0.046) and monthly income (>6 000 Yuan, β=0.085) as factors affecting occupational delay of gratification (all P<0.05). Conclusion The community healthcare workers with an education below secondary vocational school, no management position and lower income have relatively lower level of occupational delay of gratification.

Objective To explore the effect of family support service intervention on improving the rehabilitation of patients with severe mental disorders in community and the mental health status and family burden of family members. Methods Using multi-stage random sampling method, 100 patients who met the diagnostic criteria of severe mental disorders were randomly selected from two communities, and then 100 patients who met the diagnostic criteria of severe mental disorders were randomly matched according to gender, age and diagnosis in other communities into the control group. The control group and intervention group were set up strictly according to the inclusion criteria of patients and their families. Results The average age of the 200 groups was (48.27±12.67) years, and the average age of the family members was (63.61±13.19) years. After intervention, the activity dailyliving scale (ADL) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). After intervention, the social disability screening schedule (SDSS) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). After intervention, there was no significant difference in the morningside rehabilitation status scale (MRSS) score between the intervention group and the control group at all time points (all P>0.05). After intervention, the SCL-90(self-reporting inventory) scores of the mental health of the family members in the control group were higher than those in the intervention group at all times (all P<0.05). After intervention, the family burden scale of diseases (FBS) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). Conclusions The intervention measures did improve the rehabilitation effect of severe mental disorder patients in community and the psychological and family burden of family members. A professional family support service team should be established.

In recent years,a large number of studies have shown that myeloid cells in tumor microenvironment play an important role in tumorigenesis and tumor progression.On one hand,myeloid cells can regulate human immune system;on the other hand,myeloid cells can influence tumor progression,metastasis and clinical treatments.In this review,we summarize the interaction between myeloid cells and tumors,discuss the effects of myeloid cells after recruited to tumor sites and its mechanisms,try to put forward clinical therapy targeting myeloid cells and provide references for the following research and clinical treatments.

Objective To investigate the expression of DJ-1,phophatase and tensin homolog deleted on chromosome 10 (PTEN) and focal adhesion(FAK) in patients with oral squamous cell carcinoma (OSCC) and precancerous lesions.Methods Immunohistochemical staining (SP method) was used to detect the expression of DJ-1 and PTEN in the biopsy mucosa tissues from 44 cases of oral squamous cell carcinoma,20 cases of hyperplasia of oral mucosa,15 cases of dysplasia of oral mucosa and 10 cases of normal mucosa.Results The positive rate of DJ-1 in OSCC was 91％ (40/44),which was significandy higher than that in other non-malignant groups including hyperplasia(50％),dysplasia (67％)and normal mucosa(30％) (P ＜ 0.05).The positive rates of PTEN was 45％ (20/44) in OSCC,which were significantly lower than that in other non-malignant groups (P ＜ 0.05) including hyperplasia (100％),dysplasia (87％)and normal mucosa(100％) (P ＜0.05).The positive rates of FAK was 80％ (35/44) in OSCC,which were significantly lower than that in other non-malignant groups (P ＜ 0.05) including hyperplasia(100％),dysplasia(95％) and normal mucosa(100％) (P ＜ 0.05).In all of these tissues,a negative correlation was found between the expression of DJ-1 and PTEN (r =-0.75,P ＜ 0.05),while a positive correlation was found between the expression of DJ-1 and PTEN (r =0.28,P ＜ 0.05).Conclusions DJ-1,PTEN and FAK may play an important role in the pathogenesis of OSCC.

OBJECTIVETo investigate the gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time.

METHODSThe liver tissues were divided into two groups: Group A (non-hepatocirrhosis), Group B (hepatocirrhosis), each of which consisted of 3 groups with different ischemic time: 15, 30 and 45 minutes. The gene differential expression patterns in the two groups within different ischemic time were detected and compared with those in normal liver tissues by using 4000 points gene microarray.

RESULTSIn non-hepatocirrhosis tissues, the homeostatic maintenance genes expressed highly during hepatic ischemia for 15 minutes, and no apoptotic gene was expressed; but in hepatocirrhosis tissues, many apoptotic genes expressed highly. As for 30 minutes, in both two groups liver tissue genes expressed to the peak, and the genes related to cell death, oxidative stress and nuclear factors expressed highly. The difference lies in the facts that in Group B pro-apoptosis genes expressed more than those in Group A, and the Ratio values were higher than those in Group A. Many genes of heat shock protein family and antioxidant proteins expressed highly simultaneously in Group A, but comparatively low in Group B. As for 45 minutes, genes of heat shock proteins and antioxidant proteins expressed lowly in Group B.

CONCLUSIONSIt suggests that the safe time limit of hepatic ischemia for cell survive is 30 minutes or so. Non-hepatocirrhosis tissues could endure 30 minutes of ischemia and even longer, but it should be restricted within 30 minutes in hepatocirrhosis tissues.

Gene Expression Profiling ; Humans ; Ischemia ; genetics ; Liver ; blood supply ; metabolism ; Liver Cirrhosis ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis ; methods ; Time Factors

Objective: To explore the effects of microRNA-34a on regulating silent information regulator 1 (SIRT1) and influence of SIRT1 on myocardial damage of rats with severe burns at early stage. Methods: (1) Twenty-four Sprague-Dawley (SD) rats were divided into sham injury (SI) group, simple burns (SB) group and SIRT1 agonist (SA) group according to the random number table (the same grouping method below), with 8 rats in each group. Rats in groups SB and SA were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burns) on the back, and rats in group SI were sham injuried on the back. Immediately after injury, rats in groups SI and SB were intraperitoneally injected with normal saline of 50 mL/kg, and rats in group SA were intraperitoneally injected with normal saline of 50 mL/kg and 1 mg/mL resveratrol of 50 mg/kg. At 6 h post injury, abdominal aortic blood was collected to make serum and myocardial tissue of rats was collected. (2) Myocardial cells of twelve neonatal SD rats were collected and divided into microRNA-34a mimic control (MMC) group, microRNA-34a mimic (MM) group, microRNA-34a inhibitor control (MIC) group, and microRNA-34a inhibitor (MI) group, which were respectively transfected with gene sequences of mimic control, mimic, inhibitor control, and inhibitor of microRNA-34a. The microRNA-34a expression level and protein expression level of SIRT1 in myocardial cells were respectively detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Another batch of myocardial cells were divided into microRNA-34a inhibitor control+ burn serum (MCB) group, microRNA-34a inhibitor+ burn serum (MB) group, and microRNA-34a inhibitor+ burn serum + EX527 (MBE) group. Myocardial cells in group MCB were transfected with gene sequence of inhibitor control, and myocardial cells in the later groups were transfected with gene sequence of inhibitor of microRNA-34a. After transfection of 48 h, myocardial cells in group MBE were cultured in Dulbecco′s modified Eagle′s medium (DMEM) solution for 6 hours, with serum in group SB of volume fraction of 10% and final amount-of-substance concentration of 1 mol/L, and myocardial cells in the other 2 groups were cultured in DMEM solution with serum from rats of group SB of volume fraction of 10%. The protein expression levels of myocardial cells of SIRT1, cleaved-caspase-3, and Bax were detected by Western blotting. (3) Myocardial tissue from (1) was collected to detect expression levels of microRNA-34a and mRNA of SIRT1 in groups SI and SB by real-time fluorescence quantitative RT-PCR. Morphology of myocardial tissue of rats in groups SI, SB, and SA was observed with biological image navigator. The mRNA expression levels of interleukin 1β (IL-1β) and tumor necrosis factor (TNF-α) of rats in groups SI, SB, and SA were detected by real-time fluorescence quantitative RT-PCR. The expression levels of cleaved-caspase-3, and Bax of myocardial tissue of rats in groups SI, SB, and SA were detected by Western blotting. Data were processed with one-way analysis of variance and least-significant difference test. Results: (1) After transfection of 48 h, the expression level of microRNA-34a of myocardial cells in group MM was 4.67±0.92, significantly higher than 1.03±0.04 in group MMC (P<0.01); the protein expression level of SIRT1 of myocardial cells in group MM was 0.35±0.06, significantly lower than 1.12±0.11 in group MMC (P<0.01). After transfection of 48 h, the expression level of microRNA-34a of myocardial cells in group MI was 0.26±0.07, significantly lower than 1.33±0.07 in group MIC (P<0.01); the protein expression level of SIRT1 of myocardial cells in group MIC was 1.12±0.16, significantly lower than 1.74±0.34 in group MI (P<0.01). At 6 h after culture, compared with those in group MCB, the SIRT1 protein expression level of myocardial cells in group MB was significantly increased (P<0.05), while cleaved-caspase-3 and Bax protein expression levels of myocardial cells in group MB were significantly decreased (P<0.05). Compared with those in group MB, the SIRT1 protein expression level of myocardial cells in group MBE was with no significantly statistical difference (P>0.05), and cleaved-caspase-3 and Bax protein expression levels were significantly increased (P<0.05). (2) At 6 h post injury, compared with that in group SI, the microRNA-34a expression level of myocardial tissue in group SB was significantly increased (P<0.01), and the mRNA expression level of SIRT1 of myocardial tissue in group SB was significantly decreased (P<0.01). At 6 h post injury, myocardial cells in group SI arranged neatly with normal nucleus and no inflammatory cells infiltration; myocardial cells in group SB arranged disorderly, with no abnormal nucleus, and obvious inflammatory cells infiltration; myocardial cells in group SA arranged neatly, with normal nucleus and little inflammatory cells infiltration. At 6 h post injury, compared with those in group SB, the mRNA expression levels of IL-1β and TNF-α, and the protein expression levels of cleaved-caspase-3 and Bax of myocardial tissue in groups SI and SA were significantly decreased (P<0.01). Conclusions The microRNA-34a expression level of myocardial tissue of rats with severe burns at early stage increases, which decreases the expression level of SIRT1, and increases the expression levels of IL-1β, TNF-α, cleaved-caspase-3 and Bax, leading to obvious myocardial damage. Activation of SIRT1 can alleviate myocardial damage of rats with severe burns at early stage through decreasing expression levels of IL-1β, TNF-α, cleaved-caspase-3, and Bax.

Objective To compare body mass index(BMI),bioelectrical impedance analysis(BIA) and waist circumference(WC) for the assessment of the growth and development of the children,and to explore the accuracy of BIA and WC for the assessment of the growth and development of the children.Methods Children were surveyed in Department of Child Health Care of Nanjing Children's Hospital from Aug.2011 to Aug.2012.BIA was used to measure body fat,skeletal muscles and bone minerals of these children,and WC was measured by meterstick.According to BMI and body fat parameters,the study subjects were classified into lower weight,normal weight,overweight and obese groups.According to the WC,the subjects were divided into the normal group and the abnormal group.Results There were 2313 children in this survey,1528 boys of them,median age 7.7 years old(ranged from 3.0 to 16.8 years old),and 785 girls of all the children,median age 7.2 years old(ranged from 3.0 to 17.5 years old).Bone mineral content of skeletal muscles increased with age in boys and girls(P ＜0.001).According to BMI,there were 484(20.9％) obese children and 266(11.5％) lower weight children;according to the BIA,there were 663(28.7％) obese children and 481 (20.8 ％) lower weight children; according to the WC,there were 597 (25.8 ％) abnormal children.The sensitivity and specificity of WC and BIA were both higher in the evaluation of overweight and obesity status.When overweight and obesity were analyzed by BIA as well as WC,girls showed a higher specificity,while the boys showed a higher sensitivity.The sensitivity of BMI was higher than BIA in evaluating weight loss.Conclusions BIA as a commonly used indicator is reliable for population census,and it is more desirable if the measurement of WC is done at the same time.The high sensitivity of BIA makes it more suitable for screening overweight and obesity of children.