Objective To develop and verify a multiplex fluorescent quantitative PCR method for detection of common bacterial and fungal contaminants in cell culture medium.Methods According to NUC gene of Staphylococcus aureus,COLA gene of Clostridium spore,ITS-2 segment sequence of Candida albicans,a set of primers and probes were designed for each respectively,and using UBI3 gene of capsicum introduced as external standard gene,a triple reaction system of Staphylococcus aureus,Clostridium spore and external standard gene and a double reaction system of Candida albicans and external standard gene were established.The primer specificity,linear range,limit of detection,specificity,anti-interference performance and precision of the method were verified.Finally,100 samples of 293T cell culture medium were detected by using the developed method,which was compared with the common PCR method.Results Three pairs of primers all amplified about 100 bp specific gene bands corresponding to the three strains at different annealing temperatures(56,57,58 and59 ℃),and the size was consistent with the expected.In the range of 5.80 × 10~6 — 5.80 × 10~2 copies/μL,the standard plasmids of the three strains showed a good linear relationship with the Ct values.The standard curve equations were:Y=-3.373 X+37.48,Y=-3.557X+36.59 and Y=-3.536 X+39.78,each R~2> 0.99,respectively,and the amplification efficiency was in the range of 90%—110%.All the limits of detection of the three strains were 10~1 CFU/mL.The primers and probes of the three strains showed no specific amplification on the genomic DNA of six kinds of cells that were prone to cross-reaction.The genomic DNA of 293T cells,Yeast,Escherichia coli and Mycoplasma sp.had no effect on the detection.The CVs of repeatability and intermediate precision verification were both less than 15%.Among 100 cell culture medium samples,14 positive and 86 negative samples were detected,and the results of common PCR method for three positive and two negative samples randomly selected were consistent with the developed method.Conclusion The multiplex fluorescent quantitative PCR method developed in this study for the detection of bacteria and fungi in cell culture medium has good specificity,anti-interference performance and precision,and is simple to operate with low cost and high sensitivity,which can quickly detect the contaminants during cell culture.

Objective To develop and verify a multiplex fluorescent quantitative PCR method for detection of common bacterial and fungal contaminants in cell culture medium.Methods According to NUC gene of Staphylococcus aureus,COLA gene of Clostridium spore,ITS-2 segment sequence of Candida albicans,a set of primers and probes were designed for each respectively,and using UBI3 gene of capsicum introduced as external standard gene,a triple reaction system of Staphylococcus aureus,Clostridium spore and external standard gene and a double reaction system of Candida albicans and external standard gene were established.The primer specificity,linear range,limit of detection,specificity,anti-interference performance and precision of the method were verified.Finally,100 samples of 293T cell culture medium were detected by using the developed method,which was compared with the common PCR method.Results Three pairs of primers all amplified about 100 bp specific gene bands corresponding to the three strains at different annealing temperatures(56,57,58 and59 ℃),and the size was consistent with the expected.In the range of 5.80 × 10~6 — 5.80 × 10~2 copies/μL,the standard plasmids of the three strains showed a good linear relationship with the Ct values.The standard curve equations were:Y=-3.373 X+37.48,Y=-3.557X+36.59 and Y=-3.536 X+39.78,each R~2> 0.99,respectively,and the amplification efficiency was in the range of 90%—110%.All the limits of detection of the three strains were 10~1 CFU/mL.The primers and probes of the three strains showed no specific amplification on the genomic DNA of six kinds of cells that were prone to cross-reaction.The genomic DNA of 293T cells,Yeast,Escherichia coli and Mycoplasma sp.had no effect on the detection.The CVs of repeatability and intermediate precision verification were both less than 15%.Among 100 cell culture medium samples,14 positive and 86 negative samples were detected,and the results of common PCR method for three positive and two negative samples randomly selected were consistent with the developed method.Conclusion The multiplex fluorescent quantitative PCR method developed in this study for the detection of bacteria and fungi in cell culture medium has good specificity,anti-interference performance and precision,and is simple to operate with low cost and high sensitivity,which can quickly detect the contaminants during cell culture.

Objective To evaluate the feasibility of real-time triplane strain rate imaging for the quantitative assessment of left atrial function in normal subjects. Methods Totally 40 healthy volunteers were enrolled. Left atrial expansion index (LAEI), left atrial passive emptying fraction (LAPEF) and left atrial active emptying fraction (LAAEF) were measured using real-time triplane volume analysis. Mean systolic peak strain rate (MSRs), mean early diastolic peak strain rate (MSRe) and mean late diastolic peak strain rate (MSRa) were measured using real-time triplane strain rate imaging. The results were compared respectively. Results MSRs correlated positively with LAEI (r=0.61, P<0.01). MSRe correlated inversely with LAPEF (r=-0.67, P<0.01), so did MSRa with LAAEF (r=-0.78, P<0.01). Conclusion Real-time triplane strain rate imaging is feasible for the quantification of left atrial function in normal subjects.

Objective:To explore the effect of mild cognitive impairment on the subjective well-being of the elderly and the mediating role of self-efficacy in it. To provide an effective basis for health management and clinical intervention in the elderly with mild cognitive impairment.Methods:A total of 164 patients with mild cognitive impairment from December 2018 to June 2019 were surveyed by the General Self-efficacy Scale, the Montreal Cognitive Assessment Scale, and the Memorial University of Newfoundland Scale of Happiness Scale.Results:The overall score of self-efficacy was 21.41±3.98, and subjective well-being was 26.44±7.35. There was a positive correlation among cognitive function, self-efficacy and subjective well-being, except that there was no significant correlation between self-efficacy and cognitive function( r values were 0.263-0.687, P<0.01). Self-efficacy played a part in mediating between cognitive function and subjective well-being in patients with mild cognitive impairment, accounting for 37.8% of the total effect. Conclusions:Mild cognitive impairment can also have a negative impact on the subjective well-being of the elderly. Among them, self-efficacy plays a particularly important role in mediating. Interventions about self-efficacy on the elderly with mild cognitive impairment may have more significant effects.

ObjectiveTo investigate the effect of arterial stiffness on systolic deformation in hypertensive disease.MethodsSixty essential hypertensive patients were enrolled, including 25 cases with left ventricular normal geometric (group LVN) and 35 cases with left ventricular hypertrophy (group LVH) in the Affiliated Hospital of Qingdao University during July 2013 to March 2014. Thirty patients in the control group were enrolled in the same period. The peak systolic strains and strain rates were determined by using velocity vector imaging. Stroke volume was obtained by using real-time three-dimensional echocardiography. And pulse pressure/stroke volume was used as a surrogate index of arterial stiffness. Pulse pressure/stroke volume, the differences of strain and strain rate in three groups were compared by analysis of variance, and SNK-q test was used for further comparison between two groups. Multiple linear regression was performed to estimate predictors for systolic longitudinal deformation. Pearson?s correlation was used to analysis the relevance of systolic longitudinal strain and body mass index, triglyceride, left ventricular ejection fraction, age, left ventricular mass index, pulse pressure/stroke volume.ResultsPulse pressure/stroke volume were (1.26±0.45) mmHg·m2·ml-1, (1.53±0.59) mmHg·m2·ml-1, (1.82±0.43) mmHg·m2·ml-1 (1 mmHg=0.133 kPa) in the control group, LVN, LVH respectively. The systolic strains and strain rates in the control group, LVN, LVH were recorded as follows:systolic longitudinal strains were (23.60±1.94)%, (19.69±2.56)%, (17.34±2.48)%, the systolic longitudinal strain rates were (1.64±0.17) s-1, (1.52±0.14) s-1, (1.38±0.18) s-1; the systolic radial strains were (28.69±5.2)%, (30.81±4.14)%, (26.53±3.50)%, the systolic radial strain rates were (2.51±0.56) s-1, (2.60±0.45) s-1, (2.00±0.41) s-1; the circumferential strains were (24.50±5.21)%, (24.01±4.60)%, (21.00±3.70)%, the circumferential strain rates were (1.38±0.38) s-1, (1.30±0.30) s-1, (1.10±0.26) s-1. Pulse pressure/stroke volume was higher in LVN and was more pronounced in the LVH group compared with the control (LVN/LVH with the control group:q=2.90, 6.56, LVN with LVH:q=3.22, allP<0.05). The strains and strain rates in LVH were lower than those of LVN and the control group, and the differences were statistically significant. (longitudinal strains:q=15.22, 5.43; longitudinal strain rates:q=8.88, 4.54; radial strains:q=2.85, 5.36; radial strain rates:q=6.10, 6.81; circumferential strains:q=4.42, 3.61; circumferential strain rates:q=5.04, 3.42; allP<0.05). The strains and strain rates in LVN were lower than the normal group, the signiifcant differences of the longitudinal strains and longitudinal strain rates were found (q=8.73, 3.77, bothP<0.05) while there were no statistically signiifcant differences of radial strains and radial strain rates, circumferential rates and circumferential strain rates. In a multivariate analysis, LVMI and AS were found to be predictors for systolic longitudinal strain. Body mass index, triglyceride, left ventricular ejection fraction, age, left ventricular mass index and pulse pressure/stroke volume were negatively related to systolic longitudinal strain (r=-0.10,-0.09,-0.14,-0.42,-0.56, allP<0.05) by Pearson?s correlation, while LVEF was positively related to mean systolic longitudinal strain (r=0.13,P<0.05).ConclusionArterial stiffness is suitable as an predictor for left ventricular systolic deformation in hypertensive disease.

Objective: To prepare and in vitro evaluate the self-microemulsifying drug delivery system (SMEDDS) of adefovir dipivoxil (ADV).Methods: The optimized formula was screened by solubility, compatibility, ternary phase diagram and orthogonal design with the self-emulsifying time and particle size of microemulsion as the indices.The property of self-emulsification and the dissolution in vitro of ADV-SMEDDS were also determined.Results: The optimized SMEDDS was composed of Cremophor EL35 (37.5%), Transcutol HP (37.5%) and PECEOL (25%), and the drug loading was 3%.The ADV-SMEDDS formed stable microemulsion after the dilution by 50-fold amount of water in 24 s, the average particle size was (26.30±0.46) nm, the zeta potential was (-8.96±0.57) mV, and the dissolution was more than 85% in 5 min.Conclusion: The optimized formula of ADV-SMEDDS has significantly enhanced solubility and dissolution of adefovir dipivoxil in vitro.

Objective: To develop a new practical ampule cutter with simple operation. Methods: According to the actual clinical situation, the structure of the ampoule cutting device were designed, which was mainly divided into three parts: the handle, the ampoule cutting tube and the ampoule cutting device. Results: The device can effectively cut the bottleneck of ampoule bottle and avoid the injury of sharp instrument and drug contamination caused by traditional breaking the bottleneck of ampoule bottle. Conclusion This kind of ampoule cutter can break the bottleneck of ampoule effectively, avoid the cross infection of medical staff, improve the work efficiency of medical staff and the effective utilization rate of medicine.

Objective : To understand the epidemiological characteristics of notifiable respiratory infectious diseases in Huzhou City, Zhejiang Province from 2017 to 2022, so as to provide insights into formulation of respiratory infectious disease prevention and control strategies. Methods: The data pertaining to notifiable respiratory infectious disease in Huzhou City from 2017 to 2022 were collected through the Infectious Disease Report Information System of Chinese Disease Prevention and Control Information System. Epidemiological characteristics of notifiable respiratory infectious disease was analyzed using a descriptive epidemiological method. : Results Conclusions: A total of 31 314 cases of notifiable respiratory infectious diseases were reported in Huzhou City from 2017 to 2022, with an average annual reported incidence of 169.12/105. The reported incidence of notifiable respiratory infectious diseases appeared a tendency towards a rise in Huzhou City from 2017 to 2022 (P<0.05). The top six reported diseases in terms of case numbers were influenza (20 048 cases), tuberculosis (6 920 cases), COVID-19 (1 893 cases), mumps (1 413 cases), pertussis (475 cases) and scarlet fever (442 cases), accounting for 99.61% of the total cases. The incidence of influenza, COVID-19 and pertussis showed a tendency towards a rise, the incidence of mumps and tuberculosis showed a tendency towards a decline (all P<0.05), and scarlet fever remained at a low-level incidence (P>0.05). Respiratory infectious diseases were mainly reported in winter (January, February and December), with 14 644 cases accounting for 46.77%. There were 15 068 cases reported in schools and kindergartens, accounting for 48.12%. The incidence showed a U-shaped variation with age, with the highest incidence in residents at ages of 10 years and below (987.68/105), and showing a tendency towards a rise in residents at ages of 60 years and above. The incidence of respiratory infectious diseases in Huzhou City from 2017 to 2022 increased significantly. Influenza, tuberculosis, COVID-19, mumps and pertussis are key notifiable respiratory infectious diseases. Residents at ages of 10 years and below and 60 years and above should be given a high priority for respiratory infectious disease control.

Objective: To analyze the epidemiological characteristics of pertussis in Huzhou City, Zhejiang Province from 2012 to 2022, so as to provide insights into formulation of pertussis control measures. Methods: The data of reported pertussis cases in Huzhou City from 2012 to 2022 were collected through the Infectious Disease Report Information System of Chinese Disease Prevention and Control Information System. The temporal, spatial and population distributions of pertussis cases in Huzhou City from 2012 to 2022 were analyzed using a descriptive epidemiological method. Results: A total of 499 pertussis patients were reported in Huzhou City from 2012 to 2022, with mean annual reported incidence of 1.508/105, and no death was reported. The reported incidence of pertussis remained at a low level in Huzhou City from 2012 to 2021, all below 1/105, and increased to 12.625/105 in 2022. The reported incidence of pertussis appeared an overall tendency towards a rise in Huzhou City from 2012 to 2021 (Z=-29.261, P<0.001). The incidence of pertussis peaked from June to July, and a relatively higher incidence rate of pertussis was reported in Deqing (6.359/105) and Anji counties (1.725/105), while higher incidence was found among children at ages of <1 year (30.566/105), 4 years (31.896/105) and 5 years (29.485/105). Conclusion The reported incidence of pertussis was at a low level in Huzhou City from 2012 to 2021, and increased sharply in 2022. The incidence of pertussis peaked from June to July, was concentrated in Deqing and Anji counties and higher among infants under one year of age and preschool children at ages of 4 to 5 years.

Objective: To investigate the trends in incidence and epidemiological characteristics of notifiable infectious diseases in Huzhou City, Zhejiang Province from 2005 to 2023, so as to provide insights into optimizing infectious disease surveillance and control. Methods: Data of notifiable infectious disease cases in Huzhou City from 2005 to 2023 were collected from the Infectious Disease Report Information System of Chinese Disease Prevention and Control Information System. Trends in incidence were analyzed using annual percent change (APC) and average annual percent change (AAPC). The population distribution and seasonal characteristics were descriptively analyzed. Results: From 2005 to 2023, a total of 504 283 cases of notifiable infectious diseases were reported in Huzhou City, with an annual crude incidence rate of 892.65/105 and a standardized incidence rate of 989.21/105. The incidence rate of notifiable infectious diseases in Huzhou City showed an upward trend (AAPC=8.886%, P<0.05), of which there was an obvious upward trend from 2021 to 2023 (APC=95.996%, P<0.05). After the removal of COVID-19 incidence, the incidence trend was basically unchanged (AAPC=7.970%, P<0.05). From 2005 to 2023, the incidence rate of class A and B notifiable infectious diseases showed no obvious trend (P>0.05), and the incidence rate of class A and B respiratory infectious diseases showed an upward trend (AAPC=6.958%, P<0.05). After the removal of COVID-19 incidence, the two showed a downward trend (AAPC=-7.680% and -8.660%, both P<0.05). The incidence rate of class A and class B intestinal infectious diseases, blood-borne and sexually transmitted infectious diseases showed a downward trend (AAPC=-14.849% and -5.977%, both P<0.05), while the incidence rates of natural and insect-borne infectious diseases did not show a significant trend (P>0.05). The overall incidence rate of class C infectious diseases showed an upward trend (AAPC=13.058%, P<0.05). The incidence rate ratio (IRR) of notifiable infectious diseases between males and females was 1.26 (95%CI: 1.25-1.27). A total of 204 043 cases under 10 years were reported, accounting for 40.46%. The peak incidence of class A and B respiratory infectious diseases was in January, May and June, while that of class A and B intestinal infectious diseases was from June to October. The peak incidence of class C respiratory infectious diseases was in January, March, April and December, while that of class C intestinal infectious diseases was from May to August and from November to December. Conclusions The incidence rate of notifiable infectious diseases in Huzhou City showed an upward trend from 2005 to 2023, which was more obvious from 2021 to 2023. Men and children under 10 years were the high-risk population. The incidence of respiratory and intestinal infectious diseases had obvious seasonal characteristics.