Purpose To investigate the expression of protein kinase D1 (PKD1) in endometrial carcinoma and normal endometrium,and to investigate its relationship with the clinicopathological features of endometrial carcinoma.Methods Immunohistochemical SP and qRT-PCR was used to detect the mRNA and protein expression of PKD1 in 92 cases of endometrial cancer and 48 cases of normal endometrium,and to analyze the relationship of expression of PKD1 with the tumor differentiation and clinical stage of endometrial carcinoma tissue.Western blot method was used to detect the expression level of PKD1 protein in normal endometrial cell line and endometrial carcinorna cell lines with different degree of differentiation.Results mRNA and protein expression of PKD1 in endometrial carcinoma tissues was significantly higher than those in normal endometrial tissues (P ＜ 0.01),which showing a correlation to the degree of tissue differentiation and clinical pathologic staging.While,the expression level of PKD1 protein in endometrial cancer cell lines was also much higher than that in normal endometrial cells (P ＜ 0.01),and the lower differentiation,the higher level of PKD1 protein expression.Conclusion PKD1 is highly expressed in endometrial cancer patients.The level of PKD1 expression may be an important reference for predicting the malignant degree of endometrial cancer.

This paper introduces the implementation method of humanistic care nursing in the concrete work process,and puts forward some thoughts on the humanistic care nursing process,such as setting up the nursing concept with the patient-oriented;To establish the management norms of humanistic care;To develop the humanistic care model,ward or hospital,etc.In the future work will continue to deepen the connotation of humanistic care,and do a good humanistic care services.

Objective To study the application of hepatamethine cyanine near-infrared fluorescence (NIRF) dye IR-783 in the mouse models of human liver cancer exenografts, and to analyze the molecular mechanisms of the NIRF dye targeting tumor cells.Methods Luciferase-tagged HepG2 cells were inoculated subcutaneously into the nude mice.We detected the correlation of NIRF intensity and bioluminescence intensity (BIL) in the tumor region.Patient-derived xenograft (PDX) model was established in mouse by subrenal capsular implantation of clinic liver cancer specimen.After injecting the IR-783 dye, the interface between mouse kidney and the xenograft tumors was confirmed by NIRF analysis, and the tumor tissue in kidney was observed by pathology using H&E staining.The expression of CEA, AFP, HIF1α and OATP3A1 in the liver cancer tissue was detected by immunohistochemical staining.The intracellular retention of NIRF dyes was observed under fluorescence microscope after adding Mito Tracker or Lyso Tracker into cultured HepG2 cells.We added IR-783 in a co-culture system of HCCs and normal liver cells to test the specifical identification ability of IR-783 of the liver cancer cells.Results There was a good correlation between NIRF intensity and BIL intensity of the subcutaneous liver cancer xenograft region in nude mice.The margin between the mouse kidney tissue and xenograft tumors was clearly identified by IR-783.Compared with normal kidney tissue, CEA, HIF1α, OATP3A1 and AFP were highly expressed in the tumor region detected by IHC staining.The NIRF dye IR-783 was mainly accumulated in the mitochondria and lysosomes of cancer cells.GFP-tagged HepG2 cells could be recognized directly, whereas red fluorescence was not detected in normal liver cells.Conclusions IR-783 is a novel near-infrared fluorescent dye with tumor targeting and imaging properties.Its targeting ability may be related to the high expression of HIF1α and OATP3A1 in the liver cancer tissue.

According to relevant national laws and regulations, practitioner training was included into laboratory animal science teaching reform.By adjusting the training content and teaching method and use of animal models of typical human diseases, the transformation of training mode was realized and improved.By the assessment of basic theory in combination with practical operation, the thinking ability and hands-on skill of the practitioners are much improved. Through classroom instruction, experimental teaching, quality assessment and tracking survey, the evaluating process of the training quality of training teaching is performed.Therefore, the teaching reform of the laboratory animal science based on the training of practitioners is established.

Objective To study the probability of other autoimmune diseases in primary biliary cirrhosis (PBC),autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC) and explore its effects on the prognosis.Methods From January 1994 to March 2014,the data of 232 patients with autoimmune liver diseases (AILD) were collected.The type and case number of coexisting with other autoimmune diseases of patients with PBC,AIH and PSC were analyzed and compared.Cox regression model was performed to analyze the effects of coexisting with autoimmune diseases on the prognosis of AILD.Results Among 135 PBC patients,there were 64 cases that coexisted with Sjogren's syndrome (SS),seven cases with systemic lupus erythematosus (SLE),seven cases with rheumatoid arthritis (RA),nine cases with systemic sclerosis (SSc),three cases with polymyositis and/or dermatomyositis (PM/DM) and one case with Crohn's disease.Among 55 AIH patients,threre were 19 cases that coexisted with SS,10 cases with SLE,one case with RA,two cases with SSc and two cases with PM/ DM.Among 24 PSC patients,there were seven cases combined with ulceric colitis,one case with Crohn's disease and one case with RA.Among 18 patients with PBC AIH overlap syndrome,there were five cases with SS and one case with RA.Compared with PBC patients,the risk of pulmonary interstitial fibrosis increased in PBC patiento coexisting with SS (OR =34.0,95 ％ CI 8.9 to 130.1).After gender,age,disease course and medicine intervention were adjusted,the prognosis of AILD which included death,liver transplantation and liver cirrhosis complications was not affected by the coexistence with other autoimmune diseases.Conclusions AILD patients coexisting with other autoimmune diseases is common,most of which are SS,SLE,SSc and RA.PBC patients coexisting with SS is the risk factor of pulmonary interstitial fibrosis,and coexisting with other autoimmune disease does not independently affect the prognosis of AILD.

Objective To analyze monitoring data on congenital heart disease (CHD) in Shenzhen city from 2009 to 2013.Methods This study described epidemiological characteristics of perinatal CHD.Patients with CHD were divided into isolated CHD group and complex CHD group (the patients were found with malformations of other systems).The prognosis,social and demographic characteristics,and complications of pregnancy were analyzed.Results The average incidence rate of CHD was 7.25‰,of which 94.36％ were diagnosed by ultrasound,of which 33.20％ were diagnosed before delivery.The top three cardiovascular abnormalities were patent ductus arteriosus,patent foramen ovale,and ventricular septal defect.There were about 14.11％ of CHD with other system malformations,mainly including musculoskeletal system,facial features,and nervous system.Compared isolated cardiac malformations and complex cardiacmalformations,significant differences were found in the age stratification,culture degree,prognosis,diagnosis time,incidence rate of complication of pregnancy,and childbirth complications rate (P ＜0.01).Conclusions Most CHDs were diagnosed by ultrasound.Simple cardiovascular malformation was common,especially CHD with two or more types of cardiovascular malformations.The incidence rate of CHD in 30 to 40-year-old women,stillbirth rate,incidence of prenatal diagnostic rate were significantly higher than the simple CHD group.

Objective To study the tumor targeting ability and application of farnesylthiosalicylic Acid (FTS) and heptamethine carbocyanine fluorescent dye-mediated near-infrared imagine in living animals, and confirm the inhibitory effect of this compound on growth of tumor cells.Methods Human breast cancer cell line MCF-7, glioma cell line U251 and prostate cancer cell line PC3 were cultured to logarithmic growth phase, and different concentrations of FTS and FTS-IR783 were added, respectively.We observed the inhibitory effect of those two compounds on the growth of tumor cells.Under fluorescence microscopy, specific accumulation of FTS-IR783 in these tumor cells was observed.The tumor cells (1×106) were transplanted subcutaneously into nude mice.These mice were subjected to intraperitoneal injection of FTS-IR783 (10 nmol/mouse) two weeks later.In the in vivo imaging, near infrared fluorescence signal and tumor volume were measured and their correlation was analyzed.Results Compared with FTS, FTS-IR783 significantly inhibited the growth of MCF-7, U251 and PC3 cells in vitro.FTS-IR783 was specifically uptaken by these three kinds of tumor cells, showing strong near infrared fluorescence in cell agglomerates.After subcutaneous injection of FTS-IR783, the correlation between fluorescence intensity and tumor volume was 0.987, 0.998 and 0.971, respectively.Conclusions The compound of FTS conjugated with near infrared fluorescent dye IR-783 can specifically recognize tumor cells, in both in vitro and in vivo imaging.At the same time, the compound can significantly inhibit the growth of tumor cells, and may be expected to become a new potential targeted drug.

Objective To establish a patient-derived gastric cancer xenograft( PDX) model in nude mice and to in-vestigate the application of near infrared fluorescent ( NIRF) dye IR-783 in in vivo imaging of gastric cancer xenograft mod-els.Methods Fresh human gastric cancer tissue was taken and transplanted into the subrenal capsule of nude mice to es-tablish the xenograft model.When the transplanted tumors grew,took part of the tumor tissue to do HE staining and compare the structural characteristics with the primary tumor.Another portion of the tumor was xenografted into nude mice subcutane-ously.Twenty days later,the tumor-bearing mice were injected intraperitoneally with IR-783 dye (10μM) in a dose of 100 mg/20 g.The intensity of the tumor image was monitored by optical NIRF imaging.The correction between tumor volume and fluorescence intensity was analyzed.Finally,the expression of OATP1B3 and HIF1αin the xenografted tumor tissue was detected by immunohistochemistry.Results We successfully established three patient-derived xenograft ( PDH) models of human gastric cancer.The transplanted tumor tissues maintained the histological characteristics of the primary tumor well.NIRF signal can be detec ted in subrenal capsule of the xenografted nude mice.The correlation between tumor size and fluorescence intensity in the PDX models reached higher than 98%.Strong positive expressions of HIF1αand OATP1B3 in the tumor tissues were detected.Conclusions NIRF dye IR-783 can be specifically accumulated at the tumor site,therefore, can be used to detect PDX in vivo early.The tumor targeting property may be related to the expression of OATP1B3 and HIF1α.

Objective To evaluate the effect of near infrared heptamethine cyanine dye IR-783-mediated specific tumor imaging in spontaneous tumor of dogs .Methods IR-783 was intraperitoneally injected to nude mice models of transplanted tumor in a dose of 5μmol/kg.The metabolic time course of IR-783 was detected by in vivo imager .Based on the results of above observation , IR-783 was injected to dogs with spontaneous tumor in a dose of 1.5μmol/kg.The site of intravenous injection was the hind leg .Tumor and peri-tumoral tissues were removed at 5 days after IR-783-injection for fluorescence imaging , pathology and frozen section fluorescence examinations .Results After i.p.IR-783 injection to nude mice models of transplanted tumor , the transplanted tumor tissues of nude mice had stronger specific fluorescence than normal tissues by imaging at 8 days after injection .After i.v.IR-783 injection to four dogs with spontaneous tumor , the fluorescence signal in the tumor tissues was stronger than that in the normal tissues at 5 days after injection .Conclusions Near infrared fluorescent dye IR-783 could be specifically taken up by tumor tissues , and can be used for specific diagnosis of tumor.It has an important clinical application prospect .

BACKGROUND:Tumor stem cels are the root of cancer recurrence and metastasis, so clinical researches should focus on the effects of different treatments on tumor stem cels.
OBJECTIVE:To explore the effects of endocrine therapy and chemotherapy on stem cels in patients with breast cancer.
METHODS:After recovery and cultivation of estrogen receptor-positive human breast cancer cel lines MCF-7, passage 3 cels in logarithmic phase were selected and divided into three groups containing control, estradiol and estradiol with tamoxifen groups. The estradiol group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol was added into the medium, respectively; the estradiol with tamoxifen group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol with 10-6 mol/L tamoxifen were added into the medium, respectively. The same amount of absolute ethyl ethanol was added into the medium of control group. Fifteen female patients with late recurrence and metastasis of breast cancer received chemotherapy as recurrence and metastasis group. Another 15 healthy volunteers were selected as healthy control group.
RESULTS AND CONCLUSION:The proportion of CD44+CD24-/lowcel subsets in the estradiol and estradiol with tamoxifen groups was significantly higher than that of the control group (P < 0.05), and the proportion of CD44+CD24-/low cel subsets in the estradiol group was significantly higher than that of the estradiol with tamoxifen group at the same concentration (P< 0.05). The proportion of CD44+CD24-/lowcel subsets had no significant differences among groups at 10 and 20 days of culture (P < 0.05). The proportion of CD44+CD24-/low cel subsets significantly increased in MCF-7 cels after 24-hour intervention with different chemotherapy drugs. But only the proportion of CD44+CD24-/low cel subsets in the paclitaxel and doxorubicin groups was significantly higher than that of the control group after 20-day intervention (P < 0.05). Besides, the proportion of CD44+CD24-/low cel subsets in the peripheral blood of healthy volunteers was significantly lower than that of the recurrence and metastasis group (P < 0.05). Among 15 patients with late recurrence and metastatic of breast cancer, 9 had stable disease, 5 had partial remission, 1 had failed chemotherapy and cancer progression. Moreover, the proportion of CD45-CD44+CD24-/low cel subsets in the peripheral blood of patients sensitive for chemotherapy was significantly lower than that before treatment (P < 0.05). In conclusion, both endocrine therapy and chemotherapy exert a certain effect on the CD44+CD24-/low cel subsets of breast cancer positive for estrogen receptor. Given that CD44+CD24-/low cel subsets in MCF-7 cels resist chemotherapy drugs, the proportion of CD45-CD44+CD24-/low cels in the peripheral blood of patients sensitive for chemotherapy is decreased.