Objective To design a stimulating instrument for physiology experiment.Methods A sine wave was produced by a high frequency oscillator,and then the sine wave increased in voltage by a transformer to accord with the maximal range needed.Afterwards,the increscent sine wave became a direct current signal with high voltage by a rectifier and a filter.Finally,the signal became a stimulating impulse to be exported by a switch,which was controlled by a controlling signal with adjustable frequency and width.Results Voltage range adjusted between 1 and 80V;Frequency adjusted between 0.1 and 200Hz;Impulse width adjusted between 0.1 and 5ms.Conclusion The stimulating instrument designed can be used to do electricity stimulating experiment in Physiology.

Background Anti-VEGF drugs are generally applied in the treatment of ocular neovascular diseases.However,the therapy effect is unsatisfactory in some patients.Studing the effect of hypoxia-inducible factor-1 (HIF-1),a upstream regulatory gene of VEGF,and its limiting enzyme prolyl-4-hydroxylase domain proteins (PHDs) is of important clinical significance.Objective This study was to investigate the negtive regulation of exogeneous PHDs on HIF-1 pathway in human RPE cells.Methods pFLAG-PHD1,pFLAG-PHD2 and pFLAG-PHD3 plasmids were constructed by extracting RNA from Hela cell line and coloning PHD1,PHD2 and PHD3 using reverse transcription PCR with restriction enzyme.The plasmids were identified by gene sequencing.ARPE-19 cells were cultured at 21％ O2 (normoxia group),1％ O2 (hypoxia group),or in hypoxia-mimicking agents (CoCl2,anoxia group),respectively,and then were transfected with plasmids encoding FLAG-tagged PHD1,PHD2,PHD3 and pFLAGCMV2 transfected cells served as blank control.The expressional intensities of PHD1,PHD2 and PHD3 in the cells were detected and compared among different groups by using Western blot assay.The transcriptional activity of HIF-1 in the cells was evaluated with dual luciierase reporter assay.Results Western blot assay showed that PHD1,PHD2 and PHD3 all were expressed in ARPE-19 cells in the normoxia group,hypoxia group and anoxia group.The expression was strong in PHD2 protein and was weak in PHD3 protein,a statistically significant difference was found between PHD2 protein expression and PHD1 or PHD3 expressions (all at P＜0.05).Endogenous HIF-1 activity was elevated in pFLAG-CMX transfected cells in the hypoxia group and anoxia group than that in the normoxia group.Compared with pFLAG-CMX transfected cells,no obvious change was seen in the endogenous HIF-1 activity in the normoxia group,however,HIF-1 activity was declined in the hypoxia group and anoxia group after pFLAG-PHD1,pFLAG-PHD2 or pFLAG-PHD3 transfection.Under the same oxygen environment,HIF-1 activity was lower in the pFLAG-PHD2 transfected cells than that in the pFLAG-PHD1 or pFLAG-PHD3 transfected cells (both at P＜0.05).Conclusions PHDs play a negative regulation to HIF-1 activating pathway in human RPE cells,especially in hypoxia and anoxia cells.Among PHDs proteins,PHD2 presents the strongest inhibition on HIF-1 activating pathway.

Purpose　To investigate relationship between androgen receptor and apoptotic index and to further understand the tumor biology of prostate cancer. Methods　Fifty-six patients with histologically proven prostate cancer and 20 cases with benign prostatic hyperplasia(BPH) were collected. Androgen receptor(AR) were stained by 2H12 monoclonal antibody using immunohistochemical method. The apoptotic index (AI) was determined by the terminal deoxynucleotidyl transterase-mediated dUTP biotin nick end labeling (TUNEL) technique on serial sections of formalin fixed, paraffin embedded tissues. Results　AR was not found significantly difference (P>0.05) between prostate cancer and BPH. The association of AR with AI according to Gleason score were not observed in prostate cancer. AI were significantly higher in prostate cancer compared to BPH (P<0.05).AI were also significantly higher in AR-positive prostate cancer than in AR-negative prostate cancer（P<0.05）. Conclusion　AR expression can induce prostate cancer cells to become apoptosis, and may be a useful predictor in functional classification and endocrine response of prostate cancer.

Objective: To evaluate the prognostic value of chemotherapy-induced neutropenia (CIN) in metastatic colon cancer undergoing first-line chemotherapy with FOLFOX.Methods: Data were collected from a retrospective survey of 158 consecutive metastatic colon cancer patients who had undergone FOLFOX chemotherapy.The clinicopathological characteristics and chemotherapy features of the patients were analyzed as potential prognostic factors.The patients were stratified by the decreased level of CIN to three groups: large decreased level (the number of neutrophil decreased more than 1.0×109 compared with that before chemotherapy),small decreased level (the number of neutrophil decreased less than 1.0×109 compared with that before chemotherapy) and the absence of neutropenia.Results: According to a multivariate COX model, decreased level of CIN was a independent prognostic factor of colon cancer patients.Hazard ratios of death were 0.687 (95% CI: 0.381-0.812, P=0.016) for patients with large decreased level of CIN and 0.817 (95% CI: 0.527-0.939,P=0.027) for those with small decreased level of CIN compared with those of absent neutropenia patients.Median overall survival was 12.9 months (95% CI: 10.4-15.4) for patients without neutropenia (A) compared with 20.8 months (95% CI: 18.3-23.1) for patients with large-decreased level of CIN (L) and with 17.3 months (95% CI: 16.2-18.8) for those with small-decreased level of CIN (S vs.L, P=0.018;L vs.A, P=0.009;S vs.A, P=0.011).Conclusion: Our results demonstrate that the decreased level of CIN is a predictor of prognosis in patients with metastatic colon cancer undergoing FOLFOX chemotherapy.Patients who have experienced large decreased level of CIN haave longer survival time than small decreased level of CIN or absent patients.To monitor CIN decreased level timely and adjust chemotherapy drug dose may help improve the prognosis.

Objective To synthesize 68 Ga-1,4,7,10-tetraazacyclododecane-N,N',N,N()-tetraacetic acid-D-Phe1-Tyr3-Thr8-octreotide (68Ga-DOTATATE) manually and automatically,validate its qualities in vitro,and evaluate its biodistribution in ICR mice and the microPET imaging in nude mice bearing pancreatic AR42J tumor.Methods 68Ga-DOTATATE was synthesized by automatic method using commercial metal isotope multifunctional module with strong cation exchange (SCX) column and by manual method.Both the products were measured for quality control.For the biodistribution study 5 groups of ICR mice were injected with 68Ga-DOTATATE(1.11 MBq) and executed at 10,30,60,120 and 240 min postinjection,respectively.The organs were weighted,and ％ ID/g was calculated.Nude mice bearing pancreatic AR42J tumor were intravenously injected with 3.7 MBq 68Ga-DOTATATE,and then microPET imaging was acquired at 10,30,60,120,18 and 240 min.Results 68Ga-DOTATATE could be successfully synthesized by the automatic and manual methods.Both the product injections were colorless and clear.The pH value was 6.5±0.1.For the products obtained from the two methods,the radiochemical purities were over 99％,and the products were stable for 4 h at room temperature.For the automatic method,68Ga-DOTATATE was synthesized within 30 min and with the radiochemical yield of (51.8±3.2)％ (time decay corrected).For the manual method,the time used for the synthesis was 20 min,and the labeling yield was over 99％.Three batches of the products were aseptic and pyrogen-free.In ICR mice,68Ga-DOTATATE was excreted by the kidney,and showed relatively high accumulation in the liver,spleen,pancreas and adrenal glands,while lower in the bone and soft tissue.The clearance from blood was fast with (4.41±0.81) ％ID/g at 10 min postinjection and (0.78 ± 0.32) ％ ID/g at 1 h.MicroPET imaging showed increased uptake of 68GaDOTATATE in the tumor tissues,and T/NT were 2.01±0.29(10 min),6.74±2.90(30 min) and 4.46±2.05 (60 min),respectively.Conclusions 68 Ga-DOTATATE could be successfully synthesized manually and automatically.The products reach to the specification of radioactive drugs and could be used as an attractive positron emitting radiotracer for detection of the somatostatin receptor-positive tumors.

Objective:To explore the effect of quality control circle ( QQC) on the error control in PIVAS. Methods:QQC group was established in the department of PIVAS to reduce the errors in intravenous admixture practice. The status was analyzed using the total errors per week as the index, and the improvement target value was calculated by the eighty-twenty rule. The concrete causes for the errors were found out by the method of“brain storm”, and the main causes were confirmed using a fishbone diagram and the eighty-twenty rule, and then some countermeasures were summarized and carried out. The application effect of QQC was judged by the intan-gible and tangible outcomes before and after the activity, and some suggestions for the further improvement were provided. Results:Af-ter the implementation of QQC activity, the number of errors was reduced from 47 per week to 22 per week with the rate of target a-chievement of 104. 1% and the progress rate of 53. 2%. Moreover, QQC showed positive influence on the sense of being masters, co-operation ability, team spirit and sense of responsibility and confidence in the whole staff, and the ability of analyzing, summarizing and solving problems was also enhanced. Conclusion: QQC can significantly reduce the errors in the practice of intravenous admix-ture. The management method is valuable to explore and analyze the deep problems encountered in PIVAS in order to make rational and efficient measures. It is also helpful to improving the service conception of pharmacists and nurses, and enhancing the roles of pharma-cists in quality management and control to ensure medication safety.

Objective To explore the hierarchical chain management model in blood glucose control and its influence factors in patients with diabetes mellitus.Methods Health management database of diabetic patients was established in 2007 and managed by hierarchical chain management.The number of the patients reached to 1010 till 2011.The blood glucose control of diabetic patients was analyzed and its influence factors were analyzed by multivariate unconditional Logistic regression method.Results The concentration of glycosylated hemoglobin( HbA1c ) of 1010 patients with type 2 diabetes was (8.21 ±:2.70)％.Four hundred and eighty-seven cases (48.22％) reached the blood glucose standard,303 cases (30.00％)reached the blood pressure standard,245 cases (24.26％) reached the blood lipids standard,and 76 cases (7.52％) reached all three standards.Multivariate analysis showed that occupation (OR =2.521,95％ CI:1.871 - 3.397),education level (OR =1.890,95％ CI:1.642 - 2.174),disease course (OR =1.035,95％CI:1.016 -1.055),systolic pressure (OR =1.016,95％ CI:1.007 -1.025) and triglyceride (OR =1.204,95％CI:1.063 - 1.365) were the risk factors of blood glucose control (P ＜0.01).Conclusions Hierarchical chain management model is helpful for the blood glucose control in patients with type 2 diabetes.The comprehensive control and treatment of type 2 diabetes should be taken combined with related risk factors,such as blood pressure,blood lipids and diabetes disease course.

Objective To analysis the influence factors of standardization in the hierarchical chain management of type 2 diabetes and to enhance the hierarchical chain management of type 2 diabetes.Methods ( 1 ) Six hundred and ninty patients with type 2 diabetes completed 1 years management were divided into well-controlled glycosylated hemoglobin ( HbAlc ) group (＜7.0％ ) and bad-controlled glycosylated hemoglobin (HbAlc) group ( ≥ 7.0％ ).The conditions of diet,physical activity,medication,self-blood sugar monitoring and participation in health seminars were investigated and analyzed.(2) The patients were divided into standardized management group and not standardized management group.Their age,sex,educational background,occupation,monthly income per person,medical security,the course,cognition for glycuresis,two-way transfer,and chronic complications were investigated and statistically analyzed.Results ( 1 ) The proportions of physical activity (70.1％ vs 54.2％,x2=6.163,P=0.018),self-blood sugar monitoring(60.4％ vs 43.8％,x2=6.268,P=0.016) and participation in health seminars (56.0％ vs 41.7％,x2=4.577,P=0.045) in the well-controlled HbAlc group were significantly higher than those in the bad-controlled HbAlc group.(2) Their age [(61.08 ±10.04) years old vs ( 57.75 ± 9.89 ) years old,t=2.539,P=0.012],educational background ( ratio of low educational attainment:8.3 ％ vs 17.2％,x2=6.426,P=0.041 ),medical security (own expense ratios:4.6％ vs 11.5％,x2=3.543,P=0.048 ),awareness of diabetes ( ratio of poor awareness of diabetes:19.4％ vs 41.0％,x2=17.518,P=0.000 ),two-way transfer ( ratio of not transfer treatment:4.6％ vs 14.8％,x2=7.662,P=0.022) and chronic complications ( ratio of chronic complication:41.7 ％ vs 26.2％,x2=6.130,P=0.017) were significantly different between the standardized management group and not standardized management group.(3) Logistic regression analyses indicated that the age ( OR=0.954,P=0.006),monthly income per person ( OR=4.101,P=0.018 ),medical security ( OR=7.617,P=0.003 ),cognition for glycuresis ( OR=0.030,P=0.000),two-way transfer ( OR=9.079,P=0.000) and chronic complications ( OR=0.456,P=0.031 ) were the risk factors of standardized management.Conclusion We should focus on the impact factors affecting the standardized management of patients including age,monthly income per person,medical security,awareness of diabetes,ratio of not transfer treatment,positive strategies for chronic complications,improve the hierarchical chain management of type 2 diabetes,and then make the diabetic patients to early participate in standardization management of diabetes mellitus and delay the appearance of complications.

Objective To explore the hierarchical chain management model of type 2 diabetes and determine its evaluation.Method Based on the hierarchical chain management of the three community health service institutions and Dahua hospital in Shanghai Xuhui district,215 cases of type 2 diabetes had been involved in the study.Results Compared with the baseline before management,lasting blood glucose (FBG),2 h postprandial glucose (2hPBG),glycosylated hemoglobin (HbA1c),low density lipoprotein cholesterol (LDL-C),systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the diabetes after 12 months' management declined [(8.50 ±2.81) mmol/L,(11.09 ±4.01) mmol/L,(8.56 ±2.41)% ,(3.31 ± 1.06) mmol/L,(139.06 ±20.68) mm Hg (1 mm Hg = 0.133 kPa),(78.20 ± 12.11) mm Hg vs.(7.41 ±2.04) mmol/L,(9.03 ±2.46) mmol/L,(7.34 ± 1.59)% ,(3.00 ± 1.06) mmol/L,(135.48 ± 17.82) mm Hg,(77.27 ±11.83) mm Hg],and the differences were statistically significant(P＜0.01 );control rate of FBG,2hPBG,HbA1c,LDLC,SBP,DBP had improved significantly [19.5% (42/215),20.9% (45/215),24.7%(53/215),20.0%(43/215),27.4%(59/215),30.2%(65/215) vs.50.7%(109/215),53.0% (114/215),54.0%(ll6/215),42.3%(91/215),47.0%(101/215),45.6%(98/215)](P＜0.01).Conclusion Primary and secondary-care hospital based hierarchical chain management model is valid and can be implemented for type 2 diabetes.

To generate a mWAP-hLF hybrid locus that the transcription of human lactoferrin (hLF) genomic sequence is directed by the up & down stream regulatory sequence of murine whey acidic protein (mWAP) gene locus, we describe here a successive three-step 'Gap-repair' method. First, a gap-repair vector based on pBR322 vector backbone by inserting six joint homologous arms was constructed. Then using 'Gap-repair 'method mediated by Red recombination system of lambda-prophage in Escherichia coli, in the first step, the 8 kb 3' flanking region of the mWAP gene was subcloned from the Bacterial artificial chromosome which harbors the mWAP gene locus(mWAP BAC) into the gap-repair vector; in the second step, the 29 kb hLF genomic sequence from the ATG code to the TAA code was subcloned from the hLF BAC; in the third step, the 12 kb 5' flanking region of the mWAP gene was subcloned from the mWAP BAC. Finally, all these three DNA fragments were automatically combined together without any gap in the gap-repair vector, and a 49 kb mWAP-hLF hybrid locus that the hLF genomic sequence was flanked by the 5' & 3' flanking region of mWAP gene locus was constructed. The result was confirmed by PCR, restriction enzyme digestion and sequencing. Our method provide a new way for the construction of large mammary-gland expression vector.
Animals ; Bioreactors ; DNA Repair ; genetics ; Genetic Engineering ; methods ; Humans ; Hybridization, Genetic ; Lactoferrin ; genetics ; Mammary Glands, Animal ; metabolism ; Mice ; Mice, Transgenic ; Milk Proteins ; genetics