The aim of this study is to establish stable transfected cell lines which could produce SPAG4L protein labeled with Myc and His tags in vitro. The open reading frame (ORF) of human SPAG4L was amplified by PCR and the fragments were cloned into eukaryotic expression vector pcDNA3.1/myc-His(-)A. The recombined plasmids of pcDNA3.1/myc-His(-)A/SPAG4L were verified by sequencing and digestion with enzymes. Then, the recombined plasmids were introduced into HeLa cells and screened by G418. Western blotting was performed to detect the expression of SPAG4L and its tags in stable transfected cell lines. SPAG4L and its tags were expressed in the stable cell lines transfected with pcDNA3.1/myc-His(-)A/SPAG4L, but not in the control group. Further study showed that SPAG4L colocalized with the endoplasmic reticulum(ER) marker PDI by immunofluorescence. The stable transfected cell lines established in this study will provide a powerful tool for further studies such as co-immunoprecipitation and pull-down.
Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Female ; Genetic Vectors ; genetics ; HeLa Cells ; Histidine ; genetics ; Humans ; Male ; Proto-Oncogene Proteins c-myc ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Transfection

Objective:To analyze the internal conditions and external environment of high-quality development of high-level public hospitals and establish a high-quality development strategy model, for reference of public hospitals to formulate and implement high-quality development strategies.Methods:The case hospital was a pilot in high-quality development of high-level public hospitals. SWOT method was used to analyze the influencing factors, AHP analytic hierarchy process was used to compare the importance of each influencing factor, and then the four-quadrant coordinate system method was used to establish a high-quality development strategy model.Results:The total strength(S), the total weaknesses(W), the total opportunity(O), and the total threat(T)were 0.095, 0.063, 0.065 and 0.024 respectively, reflecting that the strength of internal conditions was greater than the weaknesses, and the opportunities of external environment were greater than the threats. In the four-quadrants, the growth strategy model had the largest triangle area(0.003), and the strategic quadrilateral center of gravity was located in the opportunity area of the growth strategy.Conclusions:High-level public hospitals as represented by the case hospital had entered a period of important strategic opportunities, which the external opportunities were greater than its own strength. These hospitals were recommended to fully leverage the external opportunities, adopt a positive growth oriented(SO)strategy, formulate a development strategy with their own characteristics, promote the interaction of internal conditions and external environment to form new momentum, hence achieving high-quality development.

Based on the five core indicators (the case mix index, the proportion of level-4 surgery, the proportion of technical service income in medical income, the proportion of personnel expenditure in business expenditure, and the proportion of fixed part in personnel salary), the authors discussed the guiding significance of " paying attention to the medical level, strengthening the functional orientation, reflecting the technical value, highlighting people-orientation, and ensuring sustainable development" in the high-quality development of public hospitals. In combination with the current situation of a pilot hospital of high-quality development with the corporate supports of National Health Commission and Province Government, the authors explained the common bottlenecks of public hospitals, such as inaccurate functional positioning, weak innovation ability, poor finance structure, poor salary system and insufficient attention to talents. In the last, the authors pointed out that high-quality development was the development of deepening reform, innovation driven, structural optimization, talent oriented, people-oriented and under the leadership of the Communist Party of China, and put forward targeted implementation paths to provide reference for the pilot work.

The five core indicators(the case mix index, the proportion of level-4 surgery, the proportion of technical service income in medical income, the proportion of personnel expenditure in business expenditure, and the proportion of fixed part in personnel salary) and target values determined by the pilot goal of high-quality development of high-level public hospitals jointly built by National Health Commission and Provincial Government had become the core indicators for evaluating the quality and efficiency of public hospital development. The authors proposed the optimization and improvement measures for the five core indicators one by one from the internal reform and innovation in hospitals, as well as the synergetic development and governance of " insurance-medical-medicine linkage", combined with the current situation of a pilot hospital. The authors also proposed the starting points and driving force for promoting the high-quality development of public hospitals through the synergetic development and governance of " insurance-medical-medicine linkage", including implementing government investment responsibilities, streamlining the price evaluation of medical services, deepening the reform of medical insurance payment, accelerating the expansion of drug and consumable procurement, and promoting the application of advanced technologies and products, in order to provide reference for the in-depth promotion of high-quality development of public hospitals.

With the reform of the medical and health system entering a critical period,public hospitals have also exposed new risks and challenges in economic operation.As an important means of hospital standardized management,internal control can better prevent and resolve the risk of hospital economic operation and ensure the sustainable operation of the hospital.By interpreting the requirements of current national policies on hospital internal control,it analyzes the functional positioning of financial and accounting supervision in hospital internal control,shares the internal control implementation path of sample hospitals from the perspective of financial and accounting supervision,and puts forward suggestions on strengthening internal control construction of public hospitals in the new era,in order to lay a good foundation for the high-quality development of hospitals.

Oxalate is an organic dicarboxylic acid that is a common component of plant foods.The kidneys are essential organs for oxalate excretion,but excessive oxalates may induce kidney stones.Jupiter micro-tubule associated homolog 2(JPT2)is a critical molecule in Ca2+mobilization,and its intrinsic mecha-nism in oxalate exposure and kidney stones remains unclear.This study aimed to reveal the mechanism of JPT2 in oxalate exposure and kidney stones.Genetic approaches were used to control JPT2 expression in cells and mice,and theJPT2 mechanism of action was analyzed using transcriptomics and untargeted metabolomics.The results showed that oxalate exposure triggered the upregulation of JPT2,which is involved in nicotinic acid adenine dinucleotide phosphate(NAADP)-mediated Ca2+mobilization.Tran-scriptomic analysis revealed that cell adhesion and macrophage inflammatory polarization were inhibited by JPT2 knockdown,and these were dominated by phosphatidylinositol 3-kinase(PI3K)/AKT signaling,respectively.Untargeted metabolomics indicated that JPT2 knockdown inhibited the produc-tion of succinic acid semialdehyde(SSA)in macrophages.Furthermore,JPT2 deficiency in mice inhibited kidney stones mineralization.In conclusion,this study demonstrates that oxalate exposure facilitates kidney stones by promoting crystal-cell adhesion,and modulating macrophage metabolism and in-flammatory polarization via JPT2/PI3K/AKT signaling.

Objective:To compare the differences in the detection rates of cytomegalovirus (CMV) infection between the left and right colons for guiding endoscopic biopsy-sites.Methods:The cases of colonic CMV infection confirmed by histology at the Peking Union Medical College Hospital from July 2015 to July 2018 were collected and analyzed. Splenic curvature was used as the dividing point between the left and right colons. The CMV inclusions were identified histologically, and the CMV infected cells were detected using immunohistochemistry (EnVision method). The numbers of infected cells in the left and right colons were compared by histological and immunohistochemical results. A total of 731 patients were identified. Ten of the 731 patients had histological specimens of both left and right colons and were confirmed with CMV EnVision immunohistochemical study. There were 25 biopsy or resection specimens. Seven patients were male and 3 were female, and their ages ranged from 29 to 66 years, with a median age of 55 years. All of the 10 patients were also diagnosed with ulcerative colitis.Results:The number of the cells infected with CMV in the left colon was 115 (1-41), while that in the right colon was 76 (0-51). In 8/10 cases, the number of CMV infected cells in the left colon was more than that in the right colon.Conclusions:The study on the biopsies and resection specimens suggests that CMV infection is mostly present in the left colon. The left colon thus might be an important endoscopic biopsy-site for the cases with suspicion of CMV infection.