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Objective:Build a research index performance model with discipline growth evaluation, to evaluate the scientific research performance level of various disciplines in hospital more objectively, fairly and dynamically.Methods:Take the research projects, papers, patents and achievements as the main evaluation indicators and establish the hierarchical and classified scoring standards, to form the weight index evaluation model for the total score, per capita score and growth of the discipline research.Results:Using the growth research index to evaluate the scientific research performance of departments and individual researcher between January 2018 and December 2020. Excellent departments were selected for the top 10% of the scientific research index, those whose scientific research scores increased by more than 20% compared with the previous year were selected as the progressive departments, and the top 1% of individual scientific research scores were selected as the advanced researchers, which were commended and encouraged. For the departments ranked in the bottom 5% or whose scientific research index significantly declined compared with previous year, early warning, guidance and supervision were implemented. Since the implementation of the evaluation system, the research performance of disciplines has been significantly improved, and many achievements were made.Conclusions:This evaluation mode can stimulate the enthusiasm of the disciplines and scientific researchers for entrepreneurship and innovation to set up the standards and promote the continuous improvement of the research capacity of the whole hospital.
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Scientific and technological innovation is a vital support for emergency response assurance for public health security, and it is imperative to improve the emergency-oriented research management mechanism in hospitals. In view of elements in emergency-oriented research management as well as centralized resources management and associated platform construction, the authors explored to build an emergency-oriented research model applicable to municipal hospitals, so as to better serve regional science-based epidemic prevention and elevate the capacity of scientific and technological innovation in clinical subjects.
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Objective To investigate the anesthetic efficacy of topical dyclonine hydrochloride mucilage for preputial encircling in children. Methods Sixty children under preputial encircling, 13 patients with redundant prepuce, 47 patients with phimosis, aged 4-12 years, weighing 14-38 kg, falling into ASA physical status Ⅰ or Ⅱ, were randomly divided into two groups with 30 cases each: dyclonine group (group D) and control group (group C). Children with redundant prepuce in group D were smeared evenly 1％ dyclonine hydrochloride mucilage on the anterior 2/3 foreskins, glans and coronary sulcus by anesthesiologists who were assisted by the their parents 30 min before entering the operating room. Children with phimosis in group D were smeared evenly 1％ dyclonine hydrochloride mucilage on the anterior 2/3 foreskins, and then the tube was inserted near the coronary sulcus with the 18# straight indwelling needle. The syringe was injected into the 1％ dyclonine hydrochloride mucilage, and the glans and the coronary sulcus were squeezed repeatedly several times by anesthesiologists who were assisted by the their parents 30 min before entering the operating room. The dosage of dyclonine hydrochloride mucilage for each child was 0.2-0.3 ml/kg. Children in group C were smeared evenly isodose normal saline at the same time. All the children were treated with ketamine and propofol anesthesia after entering. The occurrence of intraoperative body reaction were observed and recorded, HR and MAP were recorded before anaesthesia induction (T0), at the beginning of surgery (T1), at the time of the coronary sulcus was exposed (T2), at the time of ligating (T3), at the time of the excess foreskin was cut (T4), the dosage of ketamine and propofol were recorded, and the occurrence of postoperative recovery time and emergence agitation during recovery period were observed. Results Body dynamic reaction rate in group D was significantly lower than that in group C (P ＜ 0.05), HR and MAP was significantly lower than that in group C at T3-T4 (P ＜ 0.05), the dosage of ketamine and propofol was significantly smaller than that in group C (P ＜ 0.05), the recovery time was significantly shorter than that in group C (P ＜ 0.05), the incidence of emergence agitation was significantly decreased compared with group C (P ＜ 0.05). Conclusion Topical dyclonine hydrochloride mucilage can effectively decrease body movement, lessen cyclic fluctuation, economize general anesthetics, shorten recovery time, reduce emergence agitation in children undergoing preputial encircling.
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Objective To investigate the effects of two kinds of anesthesia methods on the immunological function of patients with primary liver cancer during perioperative period .Methods The clinical data of 86 cases of primary liver cancer from December 2010 to December 2015 were collected and divided into control group and study group ,43 cases in each group .The control group re-ceived sevoflurane inhalation anesthesia ,and the study group was given intravenous anesthesia with propofol ,the effects of different anesthesia methods on T lymphocyte subsets ,IL-2 ,IL-6 and TNF-α were compared .Results The levels of CD3+ ,CD4+ ,CD8+ , CD4+ /CD8+ ,IL-2 ,IL-6 and TNF-αin the two groups before anesthesia were not statistically significant (P>0 .05) ,there was no significant difference in CD3+ ,CD8+ and IL-2 between the two groups after anesthesia (P>0 .05) .Compared with T0(before anes-thasia) ,the two groups decreased significantly on CD4+ and CD4+ /CD8+ at the T1(end of anesthesia) segment(P<0 .05) ,but the control group returned to the pre anesthesia level in the T 2(24 h after surgery ) segment ,the indexes at the T1 and T2 segment in the study group were higher than those in the control group (P<0 .05) ,and the level of IL-6 at T1 and T2 segment were signifi-cantly higher than those at the T0 segment (P<0 .05) .Compared with T0 ,the control group increased gradually on TNF-α (P<0 .05) ,while the study group only increased at the T1 segment (P<0 .05) ,and the T2 segment was restored to the pre-anesthesia induction .Conclusion Propofol and sevoflurane could inhibit the immune function of primary hepatocellular carcinoma ,but the de-gree of inhibition of propofol is relatively light ,and with less stress response .
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Objective To investigate the effects of two kinds of anesthesia methods on the immunological function of patients with primary liver cancer during perioperative period .Methods The clinical data of 86 cases of primary liver cancer from December 2010 to December 2015 were collected and divided into control group and study group ,43 cases in each group .The control group re-ceived sevoflurane inhalation anesthesia ,and the study group was given intravenous anesthesia with propofol ,the effects of different anesthesia methods on T lymphocyte subsets ,IL-2 ,IL-6 and TNF-α were compared .Results The levels of CD3+ ,CD4+ ,CD8+ , CD4+ /CD8+ ,IL-2 ,IL-6 and TNF-αin the two groups before anesthesia were not statistically significant (P>0 .05) ,there was no significant difference in CD3+ ,CD8+ and IL-2 between the two groups after anesthesia (P>0 .05) .Compared with T0(before anes-thasia) ,the two groups decreased significantly on CD4+ and CD4+ /CD8+ at the T1(end of anesthesia) segment(P<0 .05) ,but the control group returned to the pre anesthesia level in the T 2(24 h after surgery ) segment ,the indexes at the T1 and T2 segment in the study group were higher than those in the control group (P<0 .05) ,and the level of IL-6 at T1 and T2 segment were signifi-cantly higher than those at the T0 segment (P<0 .05) .Compared with T0 ,the control group increased gradually on TNF-α (P<0 .05) ,while the study group only increased at the T1 segment (P<0 .05) ,and the T2 segment was restored to the pre-anesthesia induction .Conclusion Propofol and sevoflurane could inhibit the immune function of primary hepatocellular carcinoma ,but the de-gree of inhibition of propofol is relatively light ,and with less stress response .
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BACKGROUND:Islet beta cell replacement therapy is one of the most promising approaches for treating type 1 diabetes mellitus. However, its large scale application is hampered by a shortage of islet beta cells for transplantation. Pluripotent stem cells are one of ideal seed cells for islet beta cell replacement therapy, but pancreatic beta-cell differentiation is time-consuming and labor-intensive. OBJECTIVE:To construct a high efficient pancreatic and duodenal homeobox 1 (Pdx1)/insulin dual-reporter vector and to monitor the key genes expression during pancreatic beta-cell differentiation from pluripotent stem cells. METHODS:In order to construct a high efficient Pdx1/insulin dual-reporter vector, puromycin resistance gene was firstly introduced into pTiger vector, and then the original 410 bp mouse Ins1 promoter of the vector was replaced by 646 bp mouse Ins1 promoter. Finally, the dual-reporter vector was transduced into INS-1 and human induced pluripotent stem cells to testify its function. RESULTS AND CONCLUSION:The high efficient Pdx1/insulin dual-reporter vector was constructed successfully. The vector successfully acquired puromycin resistance gene and high gene expression efficacy of insulin in INS-1 cells. The specific gene expression pattern of Pdx1/insulin was first found in INS-1 cells. To conclude, the real-time monitoring function of Pdx1/insulin expression is preliminarily confirmed during pancreatic beta-cell differentiation.
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Objective To observe the relationship of right internal jugular vein (RIJV)and common carotid artery (CAA)by scanning strictly from the anterior to posterior using ultrasound at different head rotation.Methods Using ultrasonic scanning,the anatomic features of RIJV and CAA both at thyroid cartilage level (prominentia laryngea level)and at the apex of the angle formed by the division of the sternocleidomastoid muscle (triangle level)with 0°,1 5°,30° and 45° right rotation were observed in 131 patients with ASA physical status Ⅰ or Ⅱ (male 55 cases,female 76 cases, aged 18~74 years).Based on the ultrasound images,the safe puncture range,the overlapping ratio, the angle between the horizontal axis and the line from the midpoint of RIJV to that of CAA (αangle) were measured.In addition,the relationship between the RIJV and CAA was defined as anterior-lat-eral, lateral, posterior-lateral or extremely-posterior-lateral position according to α angle. Results The safe puncture range of RIJV augmented as head rotated from 0° to 30° position(P <0.05);The safe puncture range of RIJV at triangle level was significantly higher than at prominentia laryngea level at all the four head positions(P <0.05).The overlapping degree decreased as head rota-ted from 0°to 30°head position at prominentia laryngea level(P <0.05).No siginificant differences of the overlapping degree were found between head positons at triangle level;The overlapping degree at triangle level was less than at prominentia laryngea level when at 0°and 1 5°head positon(P <0.05). At both prominentia laryngea and triangle levels,RIJV located mainly at lateral and posterior-lateral positions.In addition,the part of lateral position increased while the part of posterior-lateral position decreased as the head rotated from 0°to 45°position(P <0.05).Conclusion The puncture conditions for RIJV catheterization were more optimal at 30°to 45°head rotation for a safer puncture range and less overlapping between RIJV and CAA.RIJV located mainly at lateral and posterior-lateral positions at different rotations and RIJV gradually shifted to lateral position while head rotation increasing.It would be much better to select triangle level for central venous catheterization than prominentia laryn-gea level.
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To establish a gene regulation system compatible with biopharmaceutical industry and gene therapy, we constructed a fusion protein of biotin ligase from Bacillus subtilis (BS-BirA) and the trans-activation domain, and used its expression vector as the regulatory vector. Meanwhile, BS-BirA-specific operators were ligated upstream of attenuated CMV promoter to obtain the response vector. In this way, a novel eukaryotic gene regulation system responsive to biotin was established and named BS-Biotin-On system. BS-Biotin-On system was further investigated with the enhancing green fluorescent protein (EGFP) as the reporter gene. The results showed that our system was superior to the current similar regulation system in its higher induction ratio, and that the expression of interest gene could be tuned in a rapid and efficient manner by changing the biotin concentrations in the cultures, Our results show that the established system may provide a new alternative for the exogenous gene modulation.
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Bacillus subtilis , Biotin , Chemistry , Eukaryotic Cells , Metabolism , Gene Expression Regulation , Genetic Vectors , Promoter Regions, Genetic , Trans-ActivatorsABSTRACT
Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
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Animals , CHO Cells , Cricetulus , Epitopes , Genetics , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Hepatitis B Vaccines , Genetics , Hepatitis B virus , Protein Precursors , Genetics , Allergy and Immunology , Recombinant Fusion Proteins , Genetics , TransfectionABSTRACT
Objective To explore the appropriate nursing care of acute hyperlipidemic pancreatitis and increase nursing level.Methods The clinical data of 3 cases of hyperlipidemic pancreatitis misdiagnosed as acute appendicitis were analyzed retrospectively and the nursing experience was summarized.Results The blood sample of all 3 cases represented dramatically high level of serum triglyceride (14.1～61.0 mmol/L).Obvious inducing factors were observed in one case.Besides upper abdominal symptom and physical sign continued in spite of the right lower abdominal discomfort,ascites was discovered by early B-ultrasound.There was no significant increase of serum or urine amylase.After correct diagnosis,all of the 3 cases recovered well by close observation,mental nursing,anti-hyperlipidemic nursing,drainage nursing and health education.Conclusions The knowledge of hyperlipidemic pancreatitis should be well understood during nursing practice.Comprehensive nursing evaluation and close observation can help doctors to analyze and estimate the disease.Integrated nursing techniques can accelerate the recovery of the patients.Health education,anti-hyperlipidemic therapy,and removal of the inducing factors are the keys of prevention.
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Taking a suspension adapted recombinant CHO cell line, 11G-S expressing human Pro-urokinase (Pro-UK) as the object of study, the impacts of different feeding nutrients, the start time of feeding and cell inoculation density on the growth and Pro-UK production of 11G-S cells in serum-free fed-batch culture were evaluated in 100 mL shacking flasks. The results indicated that amino acids, serum-free supplements and inorganic salts played important role in cell growth, cell viability and protein expression. And the effects of cells fed-batch culture was much better with the initial cell inoculation density at 3 x 10(5)-4 x 10(5) cells/mL and the start time of feeding set at 72 h, a maximum viable cells density of 7.8 x 10(6) cells/mL with a peak Pro-UK activity at 8570 IU/mL was achieved during 12 d fed-batch culture. Further, the mu of the 11G-S cells at the middle phase of the fed-batch culture, and both the q(glu) and q(gln) of the 11G-S cells at the middle and later phases of the fed-batch culture was higher than that of the 11G-S cells at the same phase of the batch culture, respectively.
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Animals , Cricetinae , Humans , CHO Cells , Cell Culture Techniques , Methods , Cricetulus , Culture Media, Serum-Free , Recombinant Proteins , Genetics , Metabolism , Urokinase-Type Plasminogen Activator , Genetics , MetabolismABSTRACT
In the light of Chinese hamster ovary (CHO) cell line 11G-S expressing human recombinant pro-urokinase, the differences of gene expression levels of the cells in different growth phases in both batch and fed-batch cultures were revealed by using gene chip technology. Then, based on the known cell cycle regulatory networks, the transcriptional profiling of the cell cycle regulatory networks of the cells in batch and fed-batch cultures was analyzed by using Genmapp software. Among the approximate 19 191 target genes in gene chip, the number of down-regulated genes was more than those of up-regulated genes of the cells in both batch and fed-batch cultures. The number of down-regulated genes of the cells in the recession phase in fed-batch culture was much more than that of the cells in batch culture. Comparative transcriptional analysis of the key cell cycle regulatory genes of the cells in both culture modes indicated that the cell proliferation and cell viability of the cells in both batch and fed-batch cultures were mainly regulated through down-regulating Cdk6, Cdk2, Cdc2a, Ccne1, Ccne2 genes of CDKs, Cyclin and CKI family and up-regulating Smad4 gene.
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Animals , Cricetinae , Humans , Batch Cell Culture Techniques , CHO Cells , Cell Cycle Proteins , Genetics , Cell Line , Cyclin-Dependent Kinase 2 , Genetics , Cyclin-Dependent Kinase 6 , Genetics , Gene Expression Profiling , Gene Expression Regulation , Recombinant Proteins , Genetics , Smad4 Protein , Genetics , Urokinase-Type Plasminogen Activator , GeneticsABSTRACT
Currently, exogenous gene expression system based on retroviral vector has been widely used as efficient gene expression system in both gene therapeutic research and RNA interference. In this study, we evaluated the efficiency of exogenous gene expression mediated by the retroviral vector in mammalian cells. First, we constructed EGFP (enhanced green fluorescent protein) vector using pcDNA3.1(+) and retroviral vector pQCXIN as backbone vector respectively. Then, we transfected or infected HEK293 cells and CHO-K1 cells with above vector or corresponding retroviral virus, and measured the relative fluorescence intensity (RFI) of EGFP. The results showed that the RFI of the retroviral virus-infected cells was two times higher than that of the plasmid-transfected cells. Further experiments revealed repeated virus infection enhanced the expression of EGFP markedly, with RFI increasing twice after four rounds of virus infection. Furthermore, the EGFP expression in HEK293 cells mediated by the retroviral vector was more stable than transfected with plasmid pcDNA3.1(+). Finally, we further validated the efficiency of exogenous gene expression system based on the retroviral vector by expressing recombinant human activated protein C (rhAPC) in HEK293 cells. We obtained HEK293 cell lines with rhAPC expression between 10 and 15 microg/(10(6) cells d). In conclusion, the exogenous gene expression system based on the retroviral vector is an alternative method for the generation of stable and high-expressing mammalian cell lines.
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Animals , Cricetinae , Humans , CHO Cells , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , HEK293 Cells , Protein C , Genetics , RNA Interference , Recombinant Proteins , Genetics , Retroviridae , Genetics , Metabolism , TransfectionABSTRACT
By using the cell density, cell viability, Pro-UK activity, specific consumption rate of glucose (q(glc)), specific production rate of lactate (q(lac)), yield of lactate to glucose (Y(lac/glc)) and as the evaluation indexes, the growth and metabolism characteristics of pro-urokinase (Pro-UK) expressing CHO cells in serum-free suspension batch culture were examined and compared to those in serum-containing suspension batch culture. We observed hardly differences in growth and metabolism characteristics between the CHO cell populations grown in serum-free suspension batch culture and serum-containing suspension batch culture. The optimal mathematical model parameters for the CHO cells grown in suspension batch culture were obtained by non-linear programming of data representing the growth, substrate consumption and product formation of the CHO cells during logarithmic growth phase using MATLAB software, and the kinetic model of the cell growth and metabolism in serum-free culture were established.
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Animals , Cricetinae , Bioreactors , CHO Cells , Cricetulus , Culture Media, Serum-Free , Culture Techniques , Methods , Kinetics , Models, Theoretical , Recombinant Proteins , Genetics , Metabolism , Urokinase-Type Plasminogen Activator , Genetics , MetabolismABSTRACT
With suspension adapted recombinant Chinese hamster ovary (CHO) cell lines 11G-S expressing human pro-urokinase (pro-UK) as the object of study, a serum-free medium for the cultivation of recombinant CHO cells in suspension was formulated by using Plackett-Burman design and response surface methodology. The two-level Plackett-Burman design was used to evaluate the effect of 10 medium supplements on the growth of the 11G-S cells in suspension culture. Among the 10 medium supplements, insulin, transferrin, and putrescine were identified as the most significant factors (P < 0.05). The response surface methodology with three factors and three levels was used to determine the optimal levels of these factors. And a serum-free medium, SFM-CHO-S for recombinant CHO cells suspension culture was formulated. The maximum cell density of 11G-S cells in SFM-CHO-S in suspension batch culture reached 4.12 x 10(6) cells/mL with a maximum pro-UK activity at 5614 IU/mL, which was superior to the commercial serum-free medium for recombinant CHO cells.
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Animals , Cricetinae , CHO Cells , Cell Culture Techniques , Methods , Cricetulus , Culture Media, Serum-Free , Genetic Engineering , Insulin , Pharmacology , Recombinant Proteins , Genetics , Transferrin , Pharmacology , Urokinase-Type Plasminogen Activator , GeneticsABSTRACT
We constructed and identified cardiac-specific a-myosin heavy chain (alpha-MHC) promoter-driven expression vector. alpha-MHC promoter was amplified by PCR by using mouse genomic DNA as template and inserted into pGEM-T Easy vector. The inserted fragment was released by enzyme digestion, and then the cytomegalovirus (CMV) promoter in pcDNA3.1(+)-EGFP-hygro vector was replaced by the alpha-MHC promoter to construct alpha-MHC-EGFP expression vector. After identification with enzyme digestion, alpha-MHC-EGFP was transfected into mouse primary cardiomyocytes by electroporation. Green fluorescence could be observed in transfected cardiomyocytes, but not in transfected non-cardiomyocytes. Alpha-MHC-EGFP expression vector was specifically expressed in cardiomyocytes, and could be used to purify embryonic stem cell-derived cardiomyocytes.
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Animals , Mice , Cell Differentiation , Cytomegalovirus , Genetics , Metabolism , DNA, Complementary , Genetics , Electroporation , Embryonic Stem Cells , Cell Biology , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Myocardium , Cell Biology , Metabolism , Myosin Heavy Chains , Genetics , Promoter Regions, Genetic , Genetics , TransfectionABSTRACT
[Objective] To analyse seminal fluid quality of seamen with unsterility,discuss the concerned environmental influencing factors.[Method] Take 128 seamen as research group,100 drivers as control;apply the Qinghua Tongfang Auto Analytic Instrument to routinly test the seminal fluid.[Result] For the seamen with unsterility,the important indexes of spermatozoon concentration,activity rate and level-a ratio were much reduced(P
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BACKGROUND: The increasing prevalence of diabetes mellitus has been become one of the diseases which threaten the heath of human being in the 21st century. Islet transplantation is considered to be the most effective approach to cure type Ⅰ diabetes mellitus. However, lack of donor tissue limits the application of this therapy. However, recent progress of stem cell research shows that stem cell therapy may be a potential means to solve this problem.OBJECTIVE: To take activin A and all-trans retinoic acid (AR) in inducing the differentiation of bone marrow mesenchymal stem cells (MSCs) and explore its possibility DESIGN: A randomized controlled experiment.SETTING: Institute of Biotechnology, Academy of Military Medical SciencesMATERIALS: This experiment was conducted at the Institute of Biotechnology, Academy of Military Medical Sciences from November 2004to June 2005. Six male Sprague-Dawley rats, with body mass of 150-160g, were provided by the Experimental Animal Center of Academy of Military Medical Sciences.METHODS: Femoral bone marrow of the rats was extracted under aseptic condition. Bone marrow mesenchymal stem cells (MSCs) were isolated with density gradient centrifugation. Passaged MSCs were randomly divided into 4 groups: high concentration of glucose (HG), AR, beta-mercaptoethanol (ME) and negative control groups. MSCs were induced to differentiate into IPCs with conditional medium containing high concentration glucose, activin A, RA and ME etc. After induction, phenotypes of differentiated cells were examined by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of insulin and glucagon of differentiated cells were examined by immunocytochemistry. Insulin-1 mR-NA expression of differentiated cells was detected by RT-PCR.RESULTS: After bone marrow mesenchymal stem cells were induced,there were scattered insulin-and glucagon-positive cells in the HG group,many insulin-and glucagon-positive cells in the AR and ME groups, and these cells formed insulin-like structure. The expression of insulin-1mRNA could be observed in the HG, AR and ME groups. Insulin-and glucagonpositive cells and the expression of insulin-1mRNA were not observed in the negative control group.CONCLUSION: We adopt an induction scheme based on AR and other matured factors, and successfully make bone marrow mesench.ymal stem cells induce and differentiate into insulin positive reaction cells and form insulin-like structure, but its induction efficiency needs further improvement.
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Objective To study the relationship between endocervical mycoplasma infection and the spontaneous abortion due to the early embryonic death and the drug sensitivity to mycoplasma. Methods Endocervical swabs were taken from fifty normal pregnant women (normal group) and fifty-eight women with spontaneous abortion due to embryonic death.The swabs were used for ureaplasma urealyticum (UU) and mycoplasma hominis (MH) Cultivation respectively. The isolation rates of the two groups were comparedf.Results (1)In the embryonic death group, the positive rates of UU, MH and UU mixed with MH were 74.1%(43/58) , 27.6% (16/58) and 20.7%(12/58)separately.In the normal female group, however, the positive rates correspondingly were the UU 48.0% (24/50), the MH 10.0% (5/50) and the UU mixed with MH 4.0% (2/50).There had significant differences of the positive rates between the two groups( P