CCK correlates with the generation and progression of pancreatic cancer. The research aims to construct eukaryotic expression plasmid pIRES2-EGFP/CCK (CCK pDNA) and transiently express it in COS-7 cells. Total RNA was extracted from porcine intestinal mucosa. RT-PCR was used to amplify the aimed segments CCKcDNA which was then digested with EcoR1 and BamH1 and inserted into a eukaryotic expression plasmid pIRES2-EGFP to construct CCK pDNA. The constructed plasmid was transfected into COS-7 cells by lepofectamin 2000-mediated transfer method. The expression of CCK in transfected COS-7 cells was detected 24, 48 and 72 h post-transfection with fluorescence microscopy and the expression level of CCK mRNA in transfected COS-7 cells was assayed by using RT-PCR. The results showed CCK pDNA was successfully constructed and expressed transiently in COS-7 cells. Green fluorescent protein could be detected in the COS-7 cells transfected with porcine CCK pDNA 24 h post-transfection. At 48th h post-transfection, the number of positive cells was increased significantly and much brighter green fluorescence could be detected. And 72 h post-transfection, the green fluorescence of positive cells became even stronger, while no green fluorescence was detected in the control group. The expression of CCK mRNA in the cells was detectable by using RT-PCR. In COS-7 cells transfected with CCK pDNA a high level of porcine CCK mRNA was detected while no expression of porcine CCKmRNA was found in the cells transfected with null plasmid. It was concluded CCK pDNA was expressed successfully in COS-7 cells, which lays a foundation for further research on the relationship between CCK and tumor.

ObjectiveTo investigate the correlation of Skp2,p27kiP1 and p21WAF1 expression with the clinicopathological features of ovarian serous cystadenocarcinomas.Methods Expressions of Skp2 ,p27kiP1 and p21WAF1 were examined by immunohistochemical staining in 124 epithelial ovarian tumors (25 serous cystadenomas, 19 borderline serous cystadenomas, and 80 serous cystadenocarcinomas) Results(1) The expression of Skp2 in serous cystadenocarcinomas (47.5％)was significantly higher than that in borderline serous cystadenomas (0％)and serous cystadenomas (0％)(P ＜ 0.001) .The p27kiP1 expression in serous cystadenocarcinomas (35.0％) was significantly lower than that in borderline serous cystadenomas(73.7％)and serous cystadenomas (80.0％) .The p21WAF1 staining frequency in serous cystadenocarcinomas (38.8％)was significantly lower than in borderline serous cystadenomas (73.7％)and serous cystadenomas (80.0％) .(2) The Skp2 protein expression in serous cystadenocarcinomas was positively correlated with clinicopathological stage,histological differentiation degree and lymph node metastasis of the tumors.The p27kiP1, p21WAF1 protein expression in serous cystadenocarcinomas was reversely correlated with clinicopathological stage and histological differentiation degree of the tumors(Ps ＜ 0.05) .(3) The Skp2 protein expression in serous cystadenocarcinomas was reversely correlated with that of p27kiP1 , p21WAF1.Conclusion The Skp2 protein expression in serous cystadenocarcinomas was increased and positively correlated with the clinicopathological features of ovarian serous cystadenocarcinomas.Skp2 protein expression was reversely correlated with p27kip1 ,p21WAF1.Skp2 protein expression may play an important role in the development and progression of serous cystadenocarcinomas.

CCK correlates with the generation and progression of pancreatic cancer. The research aims to construct eukaryotic expression plasmid pIRES2-EGFP/CCK (CCK pDNA) and transiently express it in COS-7 cells. Total RNA was extracted from porcine intestinal mucosa. RT-PCR was used to amplify the aimed segments CCKcDNA which was then digested with EcoR1 and BamH1 and inserted into a eukaryotic expression plasmid pIRES2-EGFP to construct CCK pDNA. The constructed plasmid was transfected into COS-7 cells by lepofectamine TM2000-mediated transfer method.The expression of CCK in transfected COS-7 cells was detected 24, 48 and 72 h post-transfection with fluorescence microscopy and the expression level of CCK mRNA in transfected COS-7 cells was assayed by using RT-PCR. The results showed CCK pDNA was successfully constructed and expressed transiently in COS-7 cells. Green fluorescent protein could be detected in the COS-7 cells transfected with porcine CCK pDNA 24 h post-transfection. At 48th h post-transfection, the number of positive cells was increased significantly and much brighter green fluorescence could be detected.And 72 h post-transfection, the green fluorescence of positive cells became even stronger, while no green fluorescence was detected in the control group. The expression of CCK mRNA in the cells was detectable by using RT-PCR. In COS-7 cells transfected with CCK pDNA a high level of porcine CCK mRNA was detected while no expression of porcine CCKmRNA was found in the cells transfected with null plasmid. It was concluded CCK pDNA was expressed successfully in COS-7 cells,which lays a foundation for further research on the relationship between CCK and tumor.

Objective To investigate the value of prenatal diagnosis in identifying the etiology and predicting the prognosis of fetal pleural effusion (FPE).Methods Forty-two cases of FPE were recruited in this study from January 2012 to September 2016.Ultrasound scan and genetic tests were performed on all fetuses.Seven fetuses with severe FPE were given pleurocentesis.Pregnancy outcomes of all the fetuses were followed up.Results FPE was commonly accompanied with other abnormalities,such as ascites,hydrops,hydramnion,hygroma colli,abnormal posturing,joint contractures,arrhythmia and micromandible.Chromosomal abnormality was detected in 11 fetuses (26.2％),of which ten were further confirmed by karyotype analysis,including six with 45,X,three trisomy 21 and one trisomy 18,and one was detected with a 9.83 Mb uniparental disomy (UPD) located at 12q24.21q24.31 by gene chip.One fetus was diagnosed with--SEA/--SEA thalassemia.All of the 12 families decided to terminate the pregnancies after genetic counseling.Among the other 30 fetuses,seven with severe FPE and normal karyotype underwent pleurocentesis.Five of the seven cases were with favorable outcomes,one with progressive hydrops was aborted and one neonate with severe hydrops died after birth.Spontaneous regression of FPE with good outcome was found in two cases.Parents of the other 21 fetuses chose to terminate the pregnancies.Conclusions Prenatal diagnosis is important to identify the etiology and predict the outcome of FPE.Chromosomal abnormality is a relatively common cause of FPE,and 45,X and trisomy 21 are the most common abnormalities.Intrauterine intervention is beneficial for FPE without chromosomal or other definite genetic abnormalities.Genetic test may be of great value for pregnant counseling.

Glutinous rice wine is a traditional food in south of China and it can coagulate milk. It has been proved that its function of coagulating milk is because of the presence of milk-clotting enzyme produced by microorganisms in glutinous rice wine. The aim of this work is to isolate milk-clotting enzyme producing strain from glutinous rice wine and study the fermentation condition. We screened out four bacteria and fungus by gradient dilution. It was proved that mold played the most important role in the production of milk-clotting enzyme. This is further confirmed by casein plate method. The optimization of fermentation conditions revealed that two times concentrated potato medium supplemented with 5% glucose without additional nitrogen was better for production of the enzyme. The enzyme activity was increased 144% under the conditions established.
Aspartic Acid Endopeptidases ; biosynthesis ; Bacteria ; isolation & purification ; metabolism ; Culture Media ; Fermentation ; Oryza ; metabolism ; Wine ; microbiology ; Yeasts ; cytology ; isolation & purification ; metabolism

OBJECTIVEWiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency characterized by microthrombocytopenia, eczema, recurrent infections, and an increased incidence of autoimmunity and malignancies. The patients always have a severe clinical phenotype that can result in death if not diagnosed and treated early in life. The treatment of choice with the best outcome is hematopoietic stem cell transplantation, preferably from a matched related donor. But uncertain treatment effect and high treatment cost limit its clinical application. It is the best strategy that avoiding birth of a fetus with defect through prenatal diagnosis at present. This study aimed to analyze the mutation of WASP gene in 4 Chinese families with WAS and to provide prenatal diagnosis for the high-risk fetus.

METHODThe probands of the four WAS families were all males, one of whom was deceased but had a family history and clinical datas integrated. All the patients were detected with blood routine tests, immunological tests and bone marrow examination. PCR and bilateral direct sequencing of PCR product was carried out in the regions of exon and exon-intron boundaries of WASP gene for 3 probands, 4 mothers and 100 unrelated healthy individuals as control. Prenatal diagnosis was provided for the two fetuses at the first trimester by mutation analysis.

RESULTFour WASP gene mutations were detected: c.91A > G (p.E31K), c.665C > T (p.R211X), c.397G > A (p.E133K), c.952-953delCC (p. P317fsX18), among which c.952-953delCC (p. P317fsX18) was first reported. Mothers in Family 2, 3 and 4 were carriers of WASP gene mutation, but family 1 was considered as a de-novo mutation. None of the 100 unaffected subjects had the above mutants. Prenatal diagnosis indicated that the fetus in family 2 was male and carried the same mutation as the proband, so the fetus was presumably to be a patient. The parents decided to receive an induced abortion. Following the termination of the pregnancy, the result of gene analysis of the aborted tissues was consistent with prenatal diagnosis. The fetus in family 3 was normal male confirmed by normal test results six months after birth.

CONCLUSIONThe 4 mutations of the WASP gene probably were causative to the families of WAS, among which c.952-953delCC was reported for the first time. Prenatal diagnosis by DNA sequencing is the effective method to avoid birth of WAS patient.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Fetal Diseases ; diagnosis ; Heterozygote ; Humans ; Male ; Mutation ; genetics ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis ; Sequence Analysis, DNA ; Wiskott-Aldrich Syndrome ; diagnosis ; genetics ; Wiskott-Aldrich Syndrome Protein ; genetics ; X-Linked Combined Immunodeficiency Diseases

Objective:To investigate the expressions of HIF-1α, BRD4, Beclin1, LC3B and p62 in breast cancer tissues and their clinicopathological significance, and to study alterations of their expression in breast cancer cells under hypoxic microenvironment.Methods:Immunohistochemistry was used to detect HIF-1α, BRD4, Beclin1, LC3B and p62 protein expressions in 125 breast cancer tissues and 50 para-cancer normal breast tissues, and their correlation with clinicopathologic characteristics were analyzed. The expression of these proteins were also measured after 24 hours of hypoxia stimulation was detected in different breast cancer cell lines and normal breast epithelial cells.Results:The expression of HIF-1α, BRD4, Beclin1 and LC3B proteins in breast cancer tissues were significantly higher than in para-cancer normal breast tissues ( P<0.05). There was a positive association between histologic grade, the expression of HIF-1α, BRD4, Beclin1 and LC3B ( P<0.05). High expressions of HIF-1a and Beclin1 were often correlated with lymph node metastasis and lymphatic invasion ( P<0.05). Increased HIF-1α, BRD4, Beclin1 and LC3B expression was associated with ER or PR negativity, but only HIF-1α was associated with HER2 positivity ( P<0.05). HIF-1α, BRD4, Beclin1, and LC3B were positively correlated with each other in breast cancer tissues ( P<0.01). After 24 hours of hypoxic stimulation, the expression of HIF-1α, BRD4, Beclin1 and LC3B was up-regulated in breast cancer cells. Conclusions:Hypoxia induces autophagy in breast cancer tissues. HIF-1α is positively correlated with BRD4, suggesting that BRD4 is involved in the regulation of autophagy by hypoxic microenvironment in breast cancer. High expression of HIF-1α, BRD4 and autophagy may play an important role in the development of breast cancer.

Objective:To explore effects of feedback education combined with incentive psychological intervention on compliance behavior and psychological state of patients with chronic obstructive pulmonary disease (COPD) .Methods:Using the convenient sampling method, a total of 104 COPD patients who were admitted to Xi'an Central Hospital from January 2016 to January 2019 were selected as the research objects. According to the random number table method, they were divided into the observation group and the control group, with 52 cases in each group. Patients in the control group were given conventional intervention measures, while patients in the observation group were given feedback education and incentive psychological intervention on this basis. The compliance behaviors (reasonable diet, respiratory function exercise, airway inhalation administration, rehabilitation training, disease prevention) were compared between the two group. The changes of self-care ability [Exercise of Self-care Agency Scale (ESCA) ] and mental state [Short Form of the Profile of Mood States (POMS-SF) ]before and after intervention were observed.Results:The scores of compliance with reasonable diet, respiratory function exercise, airway inhalation administration, rehabilitation training and disease prevention of patients in observation group were higher than those in control group (χ 2=4.300, 4.522, 4.833, 4.887, 4.514) . The scores of dimensions of ESCA of patients in the two groups after intervention were all higher than those before the intervention ( t=-7.189, -4.280, -8.404, -5.588, -7.51, -3.228, -8.153, -3.694) , and scores of dimensions of ESCA in the observation group were all higher than those in the control group ( t=4.819, 3.913, 5.228, 4.258) . After intervention, scores of Vigor-Activity (VA) subscale in POMS-SF of patients in the two groups were higher than those before the intervention ( t=-9.830, -8.866) , and score of the VA subscale in POMS-SF of the observation group after the intervention was higher than that of the control group ( t=5.598) . The remaining subscale scores of POMS-SF in the two groups after intervention were lower than those before intervention ( t=6.697, 6.253, 9.233, 6.612, 6.774, 5.307, 6.700, 3.096, 5.228, 2.342) , and the remaining subscale scores of the observation group after intervention were lower than those of the control group ( t=3.327, 2.675, 4.399, 4.201, 4.417) . The above differences were all statistically significant ( P<0.05) . Conclusions:During the treatment of COPD patients, feedback education and incentive psychological intervention can effectively improve compliance behaviors of patients, enhance their self-care ability, reduce their negative emotions and help them to face the disease with a positive attitude.

This article aims to provide a review of the domestic and foreign studies on the quality indicators for evaluating the psychiatric nursing care in recent years. According to Donabedian model， the research progress of quality indicators for evaluating the psychiatric nursing care were described from the three dimensions of structure quality， process quality and outcome quality. This paper summarizes the status of the research， points out the shortcomings of existing research， and provides references for the future research on the quality indicators for evaluating the psychiatric nursing care.