Social Work Intervention of Stranded Patients from the Ecosystem Perspective: Based on the Clinical Case Record Analysis of Social Work Department in Shanghai D Hospital / 中国医学伦理学

Based on the service of stranded patients in D Hospital, this study aimed to explore the multiple needs of stranded patients and construct an intervention mode from the ecosystem perspective. Using the method of case study, taking the service of stranded patients in hospital D as an example, 49 case records and verbatim interviews with stakeholders were analyzed. The discharge and resettlement of stranded patients faces multiple obstacles from individuals, families, medical and social systems. This study constructed the social work intervention model from the ecosystem perspective: information collection and needs assessment, empowerment of individual system, reconstruction of family support system, coordination of medical system and linking social support system. The intervention process achieved efficient effects. Social work intervention of stranded patients should pay attention to patients’ psychosocial needs and improve awareness of system difficulties; mobilize the multi-system support network and deliver services in combination with case management method; conduct social advocacy for the breakthrough of law and policy.

Expression of leptin and its receptor correlates with hypoxia-inducible factor-1α in infants with cyanotic congenital heart disease / 基础医学与临床

Objective To investigate the correlation between serum leptin level and body mass index(BMI)in in-fants with cyanosis congenital heart disease,and the relationship between leptin and Ob gene receptor(Ob-R)and hypoxia-inducible factor 1α(HIF-1α)in myocardium.Methods A total of 52 children under 6 months of age with congenital heart disease who underwent surgical treatment in the Department of Congenital Heart Surgery,Fuwai Hospital from January 2019 to October 2020 were included in this study.According to the arterial partial pressure of oxygen(PaO2)of 90 mmHg,they were divided into cyanotic group(n=30)and acyanotic group(n=22).Their height and weight were collected to calculate BMI.The serum leptin level was measured by ELISA.The ex-pressions of HIF-1α and Ob-R in myocardial tissue were detected by RT-PCR and Western blot.In animal mod-el,SD rats were divided into normoxia group and hypoxia intervention group,which were subjected to continuous hypoxia(10％ O2)for 4 weeks.The hypoxia intervention group received intraperitoneal injection of HIF-1α in-hibitor digoxin(2 mg/kg)daily from the 14 th to 21st day of hypoxia,respectively.The body weight of rats was recorded,and the expressions of HIF-1α and Ob-R were detected by RT-qPCR and Western blot.Results Com-pared with the acyanosis group,the cyanosis group had a significantly lower BMI(P<0.05)and a lower leptin/BMI ratio(leptin/BMI)(P<0.05).Spearman correlation analysis confirmed that serum leptin in the circulatory system was positively correlated with BMI(P<0.05).In the cyanosis group,the expression of Ob-R increased with the upregulation of HIF-1α,showing a positive correlation.In animal model,with the down-regulation of HIF-1α expression in digoxin injection,the Ob-R level was significantly lower than that in the control group(P<0.05),the trend of weight loss was significantly inhibited(P<0.05).The right ventricular hypertrophy in-dex was significantly lower than that in the control group(P<0.05).Conclusions HIF-1α regulates the expres-sion of Ob-R in myocardial tissue,and the mechanism of its association with leptin and Ob-R may help to find new therapeutic target for improving the prognosis of infants with congenital heart disease.

Research Progress in Obesity Related Heart Failure / 中国循环杂志

The prevalence rate of heart failure is increasing year by year,posing a significant threat to human health,partly due to the rapid growth of overweight and obese populations.Obesity,as an independent risk factor of heart failure,affects hemodynamics,cardiac structure and cardiac function through mechanisms such as adipocyte release of adipokines,insulin resistance and accumulation of epicardial adipose tissue.Although the results of whether weight loss can reduce heart failure incidence are inconsistent,weight loss is associated with benefits such as reduced symptoms and comorbidities,and improved quality of life.Weight loss thus plays an indispensable role in the prevention and management of cardiovascular diseases in obese individuals.Currently,effective weight loss strategies confirmed by evidence-based studies mainly include lifestyle interventions,medication therapy and weight loss surgery.

Anxiety and depression in primary and middle school students quarantined in hotels during the COVID-19 epidemic / 中国心理卫生杂志

Objective:To investigate symptoms of anxiety and depression among primary and middle school students quarantined in hotels during the COVID-19 epidemic.Methods:Anxiety and depression symptoms among 726 primary and middle school students quarantined in hotels were investigated with The Screen for Child Anxiety Related Emotional Disorders(SCARED)and Depression Self-rating Scale for Children(DSRSC)from September to October 2022 in Chifeng City,Inner Mongolia Autonomous Region.There were 624 students completed investi-gation with response rate of 86％.The positive score of SCARED was ≥23 and DSRSC was ≥ 15.Results:The detection rates of anxiety and depression were 17.9％and 15.4％respectively.The detection rates of anxiety and depression were higherin middle school students than inprimary school students(Ps＜0.05).The scores of general-ized anxiety and social phobia factors were higher in female students than in male students(Ps＜0.05).The scores of dissociative anxiety factor and depression were higher in middle school students than in primary school students(Ps＜0.05).Conclusion:During the COVID-19 epidemic,middle school students quarantined in hotels are more likely to have anxiety and depression symptoms than primary school students,and female students are more likely to have anxiety symptoms than male students.

Correlation between carotid atherosclerosis and subclinical left ventricular dysfunction in patients with type 2 diabetes / 中华内分泌代谢杂志

Objective:To investigate the relationship between carotid atherosclerosis(CAS)and subclinical left ventricular(LV)dysfunction in type 2 diabetes mellitus patients with preserved LV ejection fraction(LVEF).Methods:A total of 120 patients with type 2 diabetes mellitus who had LVEF≥50% were selected in the Department of Endocrinology, the First Affiliated Hospital of Air Force Medical University from June 2021 to October 2021. The global longitudinal strain(GLS)was obtained by two-dimensional speckle tracking echocardiography(STE)to assess subclinical LV systolic function. The mitral ratio of peak early to late diastolic filling velocity(E/A), and mitral velocity to early diastolic velocity of the mitral annulus(E/E′)ratio were obtained by pulsed tissue Doppler echocardiography to assess LV diastolic function. Acrroding to bilateral carotid ultrasound examination, the subjects were divided into normal carotid arteries group( n=46) and CAS group( n=74). Demographics and biochemical parameters were compared between two groups. Binary logistic regression and Pearson correlation analysis were used to evaluate the relationship between CAS and subclinical LV dysfunction. Results:The CAS group had a higher proportion of men, older age, and a longer duration of diabetes than the normal carotid arteries group(all P<0.05). There was no difference in LVEF and GLS between the two groups [normal carotid arteries group vs CAS group, LVEF: (60.72±4.73)% vs(60.07±4.28)%; GLS: (18.24±3.72)% vs(17.81±3.47)%, respectively; both P>0.05]. However, compared with normal carotid arteries group, E/A ratio was decreased and E/E′ ratio was significantly increased in CAS group(both P<0.01). Pearson correlation analysis showed that GLS was not correlated with carotid plaque thickness and carotid intima-media thickness(CIMT; both P>0.05). By contrast, E/E′ ratio was positively correlated with carotid plaque thickness and CIMT(both P<0.05). Binary logistic regression analysis showed that GLS and E/E′ ratio were not associated with CAS( both P>0.05). However, decreased E/A ratio was significantly associated with the existence of CAS( OR=0.09, 95% CI 0.01-0.67, P=0.018). Conclusions:In type 2 diabetes mellitus patients without overt heart failure and with preserved LVEF, the occurrence of CAS is not associated with subclinical LV systolic impairment assessed by GLS, but is significantly associated with LV diastolic dysfunction, and is independent of traditional cardiovascular risk factors.

Intervention of Traditional Chinese and Western Medicine in NLRP3 Inflammasome-mediated Digestive System Diseases： A Review / 中国实验方剂学杂志

The aberrant activation of the nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 （NLRP3） inflammasome as an essential component of the innate system is implicated in the pathogenesis of several human inflammatory diseases. Studies have confirmed its association with digestive system diseases such as ulcerative colitis， Crohn's disease， and acute pancreatitis， suggesting that the NLRP3 inflammasome plays a role in the initiation and progression of these diseases. Based on the mechanism of NLRP3 inflammasome activation and the pathways that mediate the inflammatory response， this article introduced the relationship between the NLRP3 inflammasome and the pathogenesis of multiple digestive system diseases and the Chinese and western medical therapies. Traditional Chinese medicine （TCM） has demonstrated definite effects on the NLRP3 inflammasome-mediated digestive system diseases. Some single Chinese medicines or TCM prescriptions can treat digestive system diseases by activating or inhibiting NLRP3 inflammasome activation. NLRP3 inflammasome can receive a variety of endogenous and exogenous stimulatory signals， which can initiate， activate， and mediate inflammatory responses. The inflammasome formation and downstream inflammatory cytokines are involved in not only the inflammatory responses but also the development and progression of multiple digestive system diseases. Therefore， the NLRP3 inflammasome can serve as an ideal target for disease treatment. The future rediscovery and in-depth studies of multiple inflammasomes will shed new light on the treatment of multiple digestive system diseases.

Spatial distribution of Oncomelania hupensis spread in Hubei Province from 2020 to 2022 / 中国血吸虫病防治杂志

OBJECTIVE@#To identify the spatial distribution pattern of Oncomelania hupensis spread in Hubei Province, so as to provide insights into precision O. hupensis snail control in the province.@*METHODS@#Data pertaining to emerging and reemerging snails were collected from Hubei Province from 2020 to 2022 to build a spatial database of O. hupensis snail spread. The spatial clustering of O. hupensis snail spread was identified using global and local spatial autocorrelation analyses, and the hot spots of snail spread were identified using kernel density estimation. In addition, the correlation between environments with snail spread and the distance from the Yangtze River was evaluated using nearest-neighbor analysis and Spearman correlation analysis.@*RESULTS@#O. hupensis snail spread mainly occurred along the Yangtze River and Jianghan Plain in Hubei Province from 2020 to 2022, with a total spread area of 4 320.63 hm2, including 1 230.77 hm2 emerging snail habitats and 3 089.87 hm2 reemerging snail habitats. Global spatial autocorrelation analysis showed spatial autocorrelation in the O. hupensis snail spread in Hubei Province in 2020 and 2021, appearing a spatial clustering pattern (Moran's I = 0.003 593 and 0.060 973, both P values < 0.05), and the mean density of spread snails showed spatial aggregation in Hubei Province in 2020 (Moran's I = 0.512 856, P < 0.05). Local spatial autocorrelation analysis showed that the high-high clustering areas of spread snails were mainly distributed in 50 settings of 10 counties (districts) in Hubei Province from 2020 to 2022, and the high-high clustering areas of the mean density of spread snails were predominantly found in 219 snail habitats in four counties of Jiangling, Honghu, Yangxin and Gong'an. Kernel density estimation showed that there were high-, secondary high- and medium-density hot spots in snail spread areas in Hubei Province from 2020 to 2022, which were distributed in Jingzhou District, Wuxue District, Honghu County and Huangzhou District, respectively. There were high- and medium-density hot spots in the mean density of spread snails, which were located in Jiangling County, Honghu County and Yangxin County, respectively. In addition, the snail spread areas negatively correlated with the distance from the Yangtze River (r = -0.108 9, P < 0.05).@*CONCLUSIONS@#There was spatial clustering of O. hupensis snail spread in Hubei Province from 2020 to 2022. The monitoring and control of O. hupensis snails require to be reinforced in the clustering areas, notably in inner embankments to prevent reemerging schistosomiasis.

Design and implementation of a refined management system for high value consumables in orthopedics / 中华医院管理杂志

In order to promote the high-quality development of public hospitals and ensure the safety and health of the people, it is important toachieve full traceability and refined management of high-value consumables throughout their entire life cycle.The author designed and implemented a refined management of high-value consumables in orthopedics based on one item, one code. In the closed-loop management process, hospital staff labelled high value orthopedic consumables, achieving the integration of disinfection traceability code, product qualification certificate, and billing traceability code; Three parties, including the chief surgeon, circulating nurse, and handwashing nurse, should sign and confirm the code, ensuring that the item code could not be separated; And real-time intraoperative billing, postoperative counting, and case tracking were used. At the same time, the design utilized a hospital consignment warehouse. The comprehensive promotion of this practice by the hospital saved costs, improved the overall efficiency of the entire business process of orthopedic high-value consumables, and met the requirements of relevant policy supervision and hospital refined management.

Construction of an intervention program for diabetic retinopathy patients undergoing surgery based on the theory of'Timing It Right' / 中华护理杂志

Objective To construct an intervention program for diabetic retinopathy(DR)patients undergoing surgery based on the theory of'Timing It Right'.Methods Guided by the theory of'Timing It Right',based on literature review and semi-structured interview,the first draft of the intervention program was formulated.From December 2022 to February 2023,15 medical and nursing experts in ophthalmology and endocrinology from 5 provinces(cities)in Zhejiang,Sichuan,Shanxi,Chongqing and Shanghai were interviewed by Delphi expert for 2 rounds to seek their advice and revision,and the final draft was formed.Results The effective recovery rate of 2 rounds of expert consultations was 93.75％and 100％.The authority coefficients were 0.87.The importance and feasibility of Kendall'W coordination coefficients were 0.325,0.138 and 0.193,0.141 in 2 rounds,respectively(P＜0.001).The finalized intervention program consisted of 6 months,which included 5 first-level items,18 second-level items and 43 third-level items.Conclusion The intervention program for DR patients undergoing surgery based on the theory of'Timing It Right'is reliable,scientific,feasible and practical,and it can meet the needs of such patients,providing a reference for improving their self-management ability,improving their negative emotions and delaying disease progression.

Social Work Intervention of Stranded Patients from the Ecosystem Perspective: Based on the Clinical Case Record Analysis of Social Work Department in Shanghai D Hospital / 中国医学伦理学

Based on the service of stranded patients in D Hospital, this study aimed to explore the multiple needs of stranded patients and construct an intervention mode from the ecosystem perspective. Using the method of case study, taking the service of stranded patients in hospital D as an example, 49 case records and verbatim interviews with stakeholders were analyzed. The discharge and resettlement of stranded patients faces multiple obstacles from individuals, families, medical and social systems. This study constructed the social work intervention model from the ecosystem perspective: information collection and needs assessment, empowerment of individual system, reconstruction of family support system, coordination of medical system and linking social support system. The intervention process achieved efficient effects. Social work intervention of stranded patients should pay attention to patients’ psychosocial needs and improve awareness of system difficulties; mobilize the multi-system support network and deliver services in combination with case management method; conduct social advocacy for the breakthrough of law and policy.

The relationship between insulin resistance and risk of long-term mortality in people without diabetes: a 30-year follow-up of the Daqing Diabetes Study / 中华内科杂志

Objective:To determine whether insulin resistance is associated with all-cause mortality in subjects without diabetes.Methods:A total of 505 participants without diabetes, 198 with normal glucose tolerance (NGT) and 307 with impaired glucose tolerance (IGT), were recruited from the Daqing Diabetes Study. The participants were followed up for 30 years. They were stratified into three groups (tertiles) according to baseline homeostasis model assessment of insulin resistance(HOMA-IR) levels, as the HOMA-IR 0, the HOMA-IR 1 and the HOMA-IR 2 groups, to assess the predictive effect of insulin resistance on risk of all-cause mortality.Results:During the 30-year follow-up, 52, 56 and 78 participants died across the three HOMA-IR groups, respectively. The corresponding mortality per 1 000 person-years (95 %CI) were 12.12 (9.56-15.01), 13.10 (10.46-16.03) and 19.91 (16.73-23.15), respectively. Participants in the HOMA-IR 2 group had a significantly higher risk of death than those in the HOMA-IR 0 group after adjustment of age, sex and smoking status ( HR=1.97,95 %CI 1.38-2.81, P<0.001). Cox analyses showed that a one standard deviation increase in HOMA-IR was associated with a 22% increase in the mortality after adjustment of potential confounders ( HR=1.22, 95 %CI 1.08-1.39, P=0.002). Conclusions:Insulin resistance is associated with increased risk of all-cause death in Chinese people without diabetes, suggesting that improving insulin resistance could be beneficial for people without diabetic in reducing risk of long-term all-cause mortality.

Simulation Study on the Influence of Sampling Delay on the Accuracy of Energy Metabolism Measurement / 中国医疗器械杂志

Indirect energy metabolism measurement is the gold standard for providing nutritional support for critical illness. The accuracy of the measurement data directly affects the outcome of the disease. In order to study the influence of sampling delay on the accuracy of energy metabolism measurement under mechanical ventilation, the Matlab/Simulink platform and respiratory electrical model were used for simulation and quantitative analysis. The results show that the error of indirect energy metabolism measurement increases with the increase of sampling delay, the error of sampling delay in mechanical ventilation mode is larger than that of spontaneous breathing, and the error of sampling delay in PCV mode of mechanical ventilation is larger than that in VCV mode. Therefore, there should be different sampling delay compensation strategies under severe mechanical ventilation and its different control modes.

Design and Verification of Lung Diffusion Function Detection System / 中国医疗器械杂志

A lung diffusion function detection system is designed. Firstly, the controllable collection of air, test gas source and calibration gas source was based on single-breath method measurement principle. Secondly, pulmonary diffusing capacity for carbon monoxide (DlCO) was calculated by gas concentration measured by the non-dispersive infrared sensor to measure, the gas flow measured by the differential pressure sensor, and the temperature, humidity and atmospheric pressure sensors to test and evaluate the quantitative detection and evaluation of lung diffusion function. Moreover, a preliminary verification of the lung diffusion function detection system was implemented, and the results showed that the error of the lung carbon monoxide diffusion and the alveolar volume did not exceed 5%. Therefore, the system has high accuracy and is of great value for early screening and accurate assessment of COPD.

Mechanism of action of bile acid-farnesoid X receptor-intestinal microecological axis in the development of liver failure and liver regeneration / 临床肝胆病杂志

Liver failure is a common critical medical disease, and extensive liver cell necrosis within a short period of time exceeds the regeneration capacity of liver cells and thus results in an extremely high fatality rate. Promotion of effective liver regeneration is the key to antagonizing liver failure. Recent studies have shown that bile acid, farnesoid X receptor (FXR), and intestinal microecology play an important role in liver failure and liver regeneration. This article reviews the association between bile acid, FXR, and intestinal microecology and their role in liver failure and liver regeneration, so as to provide new ideas for the treatment of liver failure in clinical practice.

Correlation between the expression of serum estradiol and otolin-1 and the recurrence of postmenopausal benign paroxysmal positional vertigo / 中国医师杂志

Objective:To explore the correlation between the expression levels of serum estradiol and otolin-1 and the recurrence of postmenopausal benign paroxysmal positional vertigo (BPPV).Methods:A total of 116 postmenopausal female patients who were diagnosed with primary BPPV in the Vertigo Treatment Center of Beijing Geriatric Hospital from January 2018 to December 2019 were selected as the research objects. They were divided into recurrence group (27 cases) and the non-recurrence group (89 cases) according to the recurrence during follow-up. The basic data, laboratory indexes and complications of the two groups were compared. The serum estradiol level was detected by electrochemiluminescence and the serum otolin-1 level was detected by enzyme-linked immunosorbent assay. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of serum estradiol and otolin-1 in the recurrence of postmenopausal BPPV patients; Multivariate logistic regression was used to analyze the risk factors of recurrence in postmenopausal BPPV patients.Results:The proportion of severe cough in the recurrence group was higher than that in the non-recurrence group ( P<0.05); the level of estradiol in the recurrence group was significantly lower than that in the non-recurrence group ( P<0.05), and the level of otolin-1 was significantly higher ( P<0.05); ROC results showed that the areas under the curve (AUCs) of serum estradiol and otolin-1 for predicting the recurrence of postmenopausal BPPV patients were 0.852 (95% CI: 0.774-0.911) and 0.722 (95% CI: 0.631-0.801) respectively, and the cut-off values were 18.09 pg/ml and 361.79 pg/ml respectively; Multivariate logistic regression analysis showed that severe cough, estradiol ≤18.09 pg/ml, and otolin-1 >361.79 pg/ml were independent risk factors for recurrence in postmenopausal BPPV patients ( P<0.05). Conclusions:The serum estradiol level of patients with postmenopausal BPPV recurrence decreases, and the level of otolin-1 increases. The abnormal level is an independent risk factor affecting the recurrence of patients with postmenopausal BPPV.

Investigation on cleaning of silicone oil attached ophthalmic surgical instruments in 22 provinces (regions,municipality) in China / 中国实用护理杂志

Objective:To investigate the cleaning status of silicone oil attached ophthalmic surgical instruments in China, so as to provide reference and suggestions for the standard cleaning process of silicone oil attached ophthalmic surgical instruments.Methods:61 hospitals in 22 provinces (regions, municipality) in China were selected as the objects of investigation. The on-site treatment, pretreatment and routine cleaning of silicone oil attached ophthalmic surgical instruments were investigated by questionnaire from November 2019 to January 2020.Results:18.033% (11/61) hospitals had mixed silicone oil attached ophthalmic surgical instruments and other instruments intraoperatively, 11.475% (7/61) hospitals didn′t wipe silicone oil and other visible contaminants postoperatively. 18.033% (11/61) hospitals didn′t carry out pretreatment and 4.000% (2/50) hospitals used saline as pretreatment solution; 54.098% (33/61) hospitals were involved in ultrasonic cleaning. 32.787% (20/61) hospitals had water temperature 31-37 ℃ and 4.918% (3/61) hospitals had water temperature>37 ℃during the cleaning process; During pretreatment and routine cleaning, 38.000% (19/50) hospitals and 54.902% (28/51) hospitals selected enzymatic detergent respectively, the contact time between instruments and detergent in specialized hospital was more standardized than that in general hospital ( P<0.001); In flushing, rinsing and final rinsing, there were 29.412% (15/51) hospitals, 11.765% (6/51) hospitals and 3.922% (2/51) hospitals used water that didn′t meet the requirements. 17.647% (9/51) hospitals, 13.725% (7/51) hospitals and 13.725% (7/51) hospitals didn′t record the time of flushing, rinsing and final rinsing. Conclusions:There are still some problems in the cleaning of silicone oil attached ophthalmic surgical instruments in specialized and general hospitals, such as no on-site treatment and pretreatment, improper control of cleaning parameters, and difficulty in ensuring water quality and water time. It is necessary to formulate the cleaning process specification of silicone oil attached ophthalmic surgical instruments as soon as possible to ensure the cleaning quality and avoid the occurrence of postoperative eye complications.

Type 2 diabetes mellitus and heart failure: Hot topic report on 70th American College of Cardiology scientific annual meeting / 中华内分泌代谢杂志

Type 2 diabetes mellitus(T2DM) and heart failure (HF) often occur concomitantly and interact with each other, leading to poor prognosis and high mortality. T2DM is also an important risk factor for HF. Based on the relevant hot topics and late-breaking clinical trials of the 70th American College of Cardiology (ACC) scientific annual meeting, this paper elaborates on the importance of diabetes management and optimal management strategy for patients with diabetes and HF from aspects of epidemiology, the relationship between T2DM and HF, cardiovascular benefits of antihyperglycemic medication and evidence-based recommendations.

Effects of Shuanghuanglian oral liquids on patients with COVID-19: a randomized, open-label, parallel-controlled, multicenter clinical trial / 医学前沿

We conducted a randomized, open-label, parallel-controlled, multicenter trial on the use of Shuanghuanglian (SHL), a traditional Chinese patent medicine, in treating cases of COVID-19. A total of 176 patients received SHL by three doses (56 in low dose, 61 in middle dose, and 59 in high dose) in addition to standard care. The control group was composed of 59 patients who received standard therapy alone. Treatment with SHL was not associated with a difference from standard care in the time to disease recovery. Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group (93.4% vs. 73.9%, P = 0.006). Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia, which was evaluated by density reduction of inflammatory focus from baseline, at day 7 (mean difference (95% CI), -46.39 (-86.83 to -5.94) HU; P = 0.025) and day 14 (mean difference (95% CI), -74.21 (-133.35 to -15.08) HU; P = 0.014). No serious adverse events occurred in the SHL groups. This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.

Optimized inverse PCR strategy for constructing multilocus mutants efficiently / 生物工程学报

Mutants of proteins are the basis for studying their structure and function, this work aimed to establish an efficient and rapid method for constructing multi-site mutants. When four or more adjacent amino acid residues need to be mutated, firstly, two long and two short primers (long primers Ⅰ/Ⅰ, short primersⅡ/Ⅱ) were designed: the long primers contain mutated sites, and the number of mutant bases is ≤20 bp, the short primers do not contain mutated sites; GC contents of the long and short primers are ≤80%, and the difference of annealing temperature is ≤40 °C. Then two sets of reverse PCR amplifications were performed using primer pairs (Ⅰ/Ⅱand Ⅰ/Ⅱ) and templates, respectively. After amplification, each system can obtain non-methylated linear plasmids which contain mutated sites, and the breakpoints of the two sets of linear plasmids amplified by primers Ⅰ/Ⅱ and Ⅲ/Ⅳ were distributed on both sides of the mutated sites. Followed by digested by DpnⅠ to remove the methylated templates, the recovered PCR products, which were mixed in an equimolar ratio, were performed another round of denaturation and annealing: the two sets of linear plasmids were denatured at 95 °C and then annealed with each other's single-stranded DNA as templates to form open-loop plasmids, and then the transformants containing the mutations will be obtained after transformed the open-loop plasmids into Escherichia coli competent cells. Results showed that, this method can mutate 4 to 11 consecutive amino acid residues (8-20 bp) simultaneously, which will greatly simplify the construction of multi-site mutants, Thereby improve the efficiency of protein structure and function research further.

Aerosol formation during non-contact "air-puff" tonometry and its significance for prevention of COVID-19 / 中华实验眼科杂志

Objective:To evaluate the aerosol and concentration (PM2.5, PM10 and aerosol particle number) formation in non-contact "air-puff" tonometry and provide suggestions for medical workers to take appropriate daily protection during the prevalence of 2019-nCoV.Methods:A cross-sectional study was carried out in this study.Thirty healthy subjects were enrolled on February 22, 2020 in Eye Hospital of Wenzhou Medical University.The intraocular pressure (IOP) was measured by non-contact "air-puff" tonometer in the ophthalmic consulting room and the hall with or without masks.Particulate matter (PM)2.5, PM10 and aerosol particles were recorded by air quality detector.The cumulative effects of IOP measurement, PM2.5, PM10 and aerosol particle number were analyzed, and the aerosol density of subjects with and without masks was compared.Results:The PM2.5, PM10 and aerosol particles produced by the non-contact "air-puff" tonometry and increased with the increase of spray times.The IOP curves of 60 eyes of 30 subjects were measured respectively in two environments of medical consulting room and medical institution hall.It was found that PM2.5, PM10 and particle number fluctuated and increased with the increase of IOP measurement person times, showing cumulative effect, and the accumulation speed of aerosol density in hall was faster than that in consulting room.The density of PM2.5 and PM10 produced without gauze mask were (53.417±2.306) and (85.350± 3.488) μg/m 3, which were higher than those of (50.567±0.862) and (80.617±1.463) μg/m 3 with gauze mask.The differences were statistically significant (P=0.028, 0.019), and significant difference was found in aerosol density between with and without gauze mask, although more aerosol was produced without gauze mask in comparison to gauze mask (P=0.123). Conclusions:Aerosol can be produced by non-contact "air-puff" tonometer spraying, and it fluctuates with the increase of spraying times, showing a cumulative effect.The aerosol and particulate matter accumulation is higher in the hall with insufficient air circulation, and more aerosol and PM can be produced without gauze mask.