Objective:To acquire the implementation effect of the neonatal Phenylketonuria(PKU)Screening Project in Dong-guan,and evaluate its economic effectiveness,so as to provide references for the prevention and control policies of genetic defects in newborns.Methods:Choosing PKU children born in Dongguan from January 2012 to December 2021(excluding BH4 deficien-cy)as the subjects,conduct the cost-benefit and cost-effectiveness analysis on the prevention and treatment of PKU.The relevant data was gained from questionnaire surveys,statistical data,medical service prices of non-profit medical institutions in Dongguan,and relevant literature materials.Results:In the past 10 years,1 245 243 newboms were screened for PKU in Dongguan,the average screening rate is 93.32％.Among them,29 were diagnosed with PKU,with a positive rate of 1:42939.During continuous follow-up,23 cases were diagnosed.Among the 23 follow-up cases,only 1 had intelligence at the critical value,while the rest were normal.The total cost of screening is 29.728 7 million,and the total benefit is 14.239 9 million.The cost-effectiveness ratio is 102.51:1,the cost-benefit ratio is 1:4.79.Conclusion:Neonatal phenylketonuria(PKU)screening in Dongguan has significant social and economic benefits.It's necessary to continue and improve the screening work,especially the policy which can benefit the PKU's full life.

Cordycepin,which has great immunomodulatory activities such as anticancer,antifungal,antivirus,antileukemia and lipid-lowering ones,is the secondary metabolite of Cordyceps militaris (C.militaris).Liquid submerged fermentation is the common cultivation process to produce cordycepin.To optimize the fermentation process and improve production,monitoring the cordycepin secretion in the fermen-tation is essential.The measurement based on chromatography-mass spectrometry methods is generally involved in the complex sample pretreatments and time-consuming separation,so more rapid and convenient methods are required.Matrix-assisted laser desorption ionization mass spectrometry(MALDI-MS) is more attractive for faster and direct detection.Therefore,MALDI-MS detection combined with isotope-labeled internal standard was applied to the measurement of cordycepin content in the fermentation broth and mycelium.This method made accurate quantification of cordycepin in the range of 5-400 μg/mL with a relative standard deviation of 5.6％.The recovery rates of fermentation samples after the 1,13,and 25 days were 90.15％,94.27％,and 95.06％,respectively.The contents of cordycepin in the mycelium and fermentation broth were 136 mg/g and 148.39 mg/mL on the 20th culture day,respectively.The cordycepin secretion curve of the liquid fermentation of C.militaris was real-time traced over 25 days.

Objective: To investigate the dynamic changes of antibodies induced by leptospiral vaccines. Methods: Antigens for antibody detection were screened out. ELISA was used to analyze antibody responses induced at different time points after immunizing guinea pigs with different batches of leptospiral vaccines from different manufacturers. To investigate the relationship between antibody responses induced by leptospiral vaccines and their protective effects in animal model, guinea pigs were challenged with Leptospira after immunization. Results: There was no significant antigen-antibody reaction between the LigA protein or Patoc Ⅰ antigen and the serum samples of guinea pigs immunized with leptospiral vaccines. Notable IgG and IgM antibody reactions were observed in all vaccination groups when using bacterial proteins from seven Leptospira reference strains which were used for the preparation of leptospiral vaccines as envelope antigens. Antigen-specific IgG antibodies peaked at 35 d after the last immunization, and the highest peak of antigen-specific IgM antibodies was reached 11 d after the last immunization. Results of the challenge test showed that non-diluted leptospiral vaccines induced significant IgG and IgM antibody reactions in guinea pigs as compared with those diluted three or nine times, showing good protective effects. Conclusions Analysis of the dynamic changes of antibodies induced by leptospiral vaccines revealed that there was correlation between the induced serum antibody responses and the protective effects. This study provided reference for further study on alternative methods for evaluating leptospiral vaccine potency.

Objective To compare the prognosis of vascular in situ and bridge vessel percutaneous coronary intervention ( PCI) therapy strategies in patients with recurrent angina after coronary artery bypass grafting ( CABG) . Methods A total of one hundred and two patients with recurrent angina after CABG from January 2008 to January 2016 were involved in this study and were divided into two groups according to interventional therapy strategy:74 patients in the vascular in situ PCI group ( in situ group,74 cases) and 28 patients for bridge vessel PCI group ( bridge vessel group,28 cases) . The patients have been followed up for (33. 6± 10. 2) months. The major adverse cardiovascular events ( MACE) of the two groups were recorded, including non?fatal acute myocardial infarction ( AMI) ,target vessel revascularization ( TVR) and cardiac death, and multivariate logistic regression analysis was used to analyze the related factors of MACE. Results Compared with the bridge vessel group,the non?MACE survival rate,non?AMI survival rate and non?TVR survival rate of the in situ group were significantly increased ( ( 71. 6% ( 53/74 ) vs. 57. 1% ( 16/28 ) , 93. 2% ( 69/74 ) vs. 82. 1% (23/28),81. 1% (60/74) vs. 67. 9% (19/28) ),the differences were statistically significant (χ2=8. 141,4. 219,5. 436, P<0. 05) . Multivariable logistic regression analysis showed that age of bridge ( OR=1. 023,95%CI 1. 005-1. 026,P=0. 019) ,diabetes mellitus ( OR=2. 386,95%CI 1. 425-3. 991,P=0. 003) and bridge vessel PCI (OR=1. 884,95%CI 1. 093-3. 220,P=0. 025) were factors that affect the clinical prognosis in patients with recurrent angina pectoris after CABG. Conclusion The clinical prognosis of the in situ PCI is better than bridge vascular PCI in patients with recurrent angina after CABG,while the age of bridge, diabetes mellitus, vascular interventional treatment are factors for the effect of interventional therapy patients prognosis. The clinical prognosis is much better in native vessel PCI than that of bridge vessel PCI in patients with recurrent angina after CABG. The age of bridge,diabetes mellitus and bridge vessel PCI are the factors that affect the clinical prognosis in the patients.

Objective To explore the mechanism by which tumor necrosis factor alpha(TNF-α) induces RIP1 kinase-dependented apoptosis in L929-A fibroblastoma cells.Methods The sub-mitochondrial localization of receptor-interacting protein 1(RIP1),caspase-8 and Bid proteins was detected by dose-gradient trypsin digestion and Western blotting.The levels of reactive oxygen species (ROS),intracellular calcium concentration,mitochondrial membrane potential (MMP),and cellular adenosine triphosphate(ATP) content were determined by fluorescent probe labeling and flow cytometry assay.The mitochondrial respiratory chain complex Ⅰ and Ⅲ activities were detected by commercial kits.Nec-1,A RIP1 kinase specific inhibitor,and RIP1-/-or Bid-/-L929-A cells were used to detect the roles of RIP1 kinase and Bid protein in cell death.Results RIP1,caspase-8 and Bid proteins were co-located in the outer membrane of mitochondrial.TNF-α exposure for 3 h could induce Bid cleavage,inhibit mitochondrial respiratory chain complex Ⅲ activity and reduce MMP.Following these changes and after TNF-α exposure for 6-12 h,the intracellular calcium concentration and ROS were increased,whereas the ATP concentration was decreased,and the cells were killed.Inhibiting RIP1 kinase or knockdown RIP1 or Bid protein could suppress all the cytotoxic effects of TNF-α.Conclusion TNF-α treatment can result in RIP1 kinase-mediated Bid cleavage and inhibit mitochondrial respiratory chains and cell energy metabolism,which ultimately leads to the death of L929-A cells.

Objective To compare the efficacy of native vessel percutaneous coronary intervention (NV-PCI) and optimal drug therapy (ODT) in patients with recurrent angina after coronary artery bypass grafting (CABG). Methods The clinical data of 142 recurrent angina pectoris after CABG patients who had underwent coronary angiography were retrospectively analyzed. Among the patients, 70 cases were treated with NV-PCI (NV-PCI group), and 72 cases were treated with ODT (ODT group). The incidence of major adverse coronary events (MACE) and left ventricular ejection fraction (LVEF) were compared between 2 groups. Results All patients were followed up for at least 1 years. There were no statistical differences in the number of bypass vessels and number of occluded vessels between ODT group and NV-PCI group: (2.5 ± 0.7) branches/case vs. (2.4 ± 0.9) branches/case and (1.4 ± 0.9) branches/case vs. (1.3 ± 0.7) branches/case, P>0.05. The incidence of MACE in NV-PCI group was significantly lower than that in ODT group: 12.9％ (9/70) vs. 22.2％ (16/72), and the LVEF was significantly higher than that in ODT group:(63.5 ± 14.0)％vs. (57.1 ± 9.0)％, and there were statistical differences (P<0.05). Conclusions Compared with the ODT, the NV-PCI has lower incidence of MACE and higher LVEF in patients with recurrent angina pectoris after CABG.

Objective To explore the influence of candesartan (an angiotensin II receptor 1 antagonist,AT1R) in radioresistance of human nasopharyngeal carcinoma CNE1 cells.Methods Cell growth of CNE1 with or without candesartan treatment was measured in vitro by MTT method;radiosensitivity of CNE1 with or without candesartan treatment was tested under normoxic or hypoxic conditions by clone formation assay.The expression of hypoxia-induced factor 1α(HIF-1α)in CNE1 cells was analysed by western blotting.Results Candesartan did not significantly inhibit the growth of CNE 1 cells in both normoxic and hypoxic conditions.Candesartan also did not influence the radiosensitivity of CNE1 cells in normoxic condition;however,it significantly increased the radiosensitivity of CNE1 cells in hypoxic condition.The expression of hypoxia-induced factor 1 α (HIF-1 α)in hypoxic CNE1 cells was significantly inhibited by candesartan treatment.Conclusion Candesartan does not significantly influence the proliferation of CNE1 cells in both normoxic and bypoxic conditions but significantly enhances the radiosensitivity of hypoxic CNE1 cells,in which the mechanisn may be involved in its inhibiting HIF1α expression in hypoxic CNE1 cells.

Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.

Aim To study the influence of Sodium fer-ulate ( SF) on bone metabolism in glucocorticoid–in-duced osteoporosis rats. Methods Thirty cases of fe-male Wistar Rats(3-month-old) were divided into con-trol group, model group and SF group ( low-dose group, middle-dose group, high-dose group ) by ran-domized block design. Double fluorochrome labeling with calcein was performed before necropsy. The left tibia was taken for bone histomorphometry. Results In static parameters, the proximal tibia cancellous bone trabecular thickness, trabecular quantity and area ratio were significantly reduced in model group compared with control group;while compared with model group, those were increased in middle and high-dose SF group. Trabecular separation degree was increased in model group compared with control group, while it was decreased in middle and high-dose SF group compared with model group. In dynamic parameters, the calcula-tion parameters of cancellous bone mark perimeter rate and the bone formation rate were increased in model group compared with control group, in middle and high-dose SF group the bone formation rate was in-creased compared with model group. In bone cells, os-teoclast number per mm, osteoblast number per mm, percent osteoblast surface perimeter and percent osteo-clast surface perimeter were increased in model group compared with control group. In growth-plate, the thickness of growth-plate was increased in model group compared with control group. In bone cells and growth-plate there was no statistical significance between treat-ment group and model group. Conclusion This study demonstrates that SF can increase bone mass and im-prove bone structure,which may be related to the im-provement of bone formation. SF is effective for GIOP in rats.

Objective To analyze on the responsibility of government for drug price addition canceling policy on incomes . Methods We Collected the 3-A hospital financial information statements and drug use data from January 2013 to September 2013 ( before reform) and from October 2013 to June 2014 ( after reform) .Results After taking evaluation of drug price addition cance-ling policy on incomes in the hospital, outpatient visits grew 9.62%, discharges increases 3.50%, and the outpatient average com-pensation fell by 4.69% and hospitalization fees per person increase 3.58%.Conclusion Preliminary results were obtained in con-trolling health care costs, medical services in ascension.The government's economic compensation economic compensation is difficult to make up for the cancellation of drug addition and reduce the reasonable income .It is suggested that we should improve the compen-sation scheme and assessment standards, and further strengthen the hospital internal management.