1.Construction and characterization of recombinant human coagulation factor Ⅶ stable transfected cell lines
Xiaoxiao LI ; Jiabin CHEN ; Jiajun LIU ; Zhifei ZHANG ; Sen ZOU ; Lihua ZHU ; Zhaoyong YANG
Acta Universitatis Medicinalis Anhui 2026;61(1):16-22
ObjectiveTo construct a stable monoclonal human embryonic kidney 293 (HEK293) cell line expressing recombinant human coagulation factor Ⅶ (rhFⅦ) and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells, and monoclonal stable transfected cell lines were selected using geneticin (G418). The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction (RT-PCR). The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay (ELISA), and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence, indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8, and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was (1.27±0.09) mg/L, and the highest procoagulant activity was (380.29±13.80)%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.
2.Effects of cough enhancement techniques on weaning in adult patients under mechanical ventilation:a systematic review
Qian CHEN ; Junjun ZOU ; Chunyang XU ; Lihua GU ; Xiaojuan JIANG ; Mengqian GU
Modern Clinical Nursing 2025;24(4):81-91
Objective To systematically evaluate the impact of cough enhancement techniques on weaning outcomes in adult patients under mechanical ventilation(MV).Methods Databases from the inception to 15th November,2024 were searched for randomised controlled trials(RCTs)on cough enhancement technique in MV patients.The searched databases were CNKI,Wanfang Data base,SinoMed,Vip,PubMed,Web of Science,Cochrane Library,Embase,and CINAHL.Literature and evidence quality were evaluated using Cochrane's evaluation tool for intervention research and the GRADE system,and data were extracted and analysed via Meta-analysis with Review Manager 5.3.Results A total of 27 studies were included,involving 2,322 patients(1,169 in the trial group and 1,153 in the control group).Compared with the control group,the trial group had shorter time of mechanical ventilation[MD=-1.40,95%CI(-1.83,-0.97),P<0.001],lower weaning failure rate[OR=0.48,95%CI(0.27,0.85),P=0.01],higher oxygenation index[MD=35.48,95%CI(21.53,49.43),P<0.001],lower pulmonary atelectasis incidence[OR=0.30,95%CI(0.18,0.49),P<0.001],greater sputum volume[MD=1.88,95%CI(0.76,3.01),P=0.001],shorter ICU stay[MD=-1.90,95%CI(-2.95,-0.85),P<0.001]and lower serum procalcitonin(PCT)[MD=-0.89,95%CI(-1.64,-0.14),P=0.02],all with statistically significant differences.However,there was no significant difference in the ventilator-associated pneumonia(VAP)between the two groups[OR=0.54,95%CI(0.21,1.35),P=0.19].Conclusion Cough enhancement techniques promote sputum discharge,improve infection and oxygenation,shorten mechanical ventilation and ICU stay,reduce weaning failure and atelectasis rates in MV patients.Further studies are required to reduce the incidence of VAP.
3.Pulmonary hamartoma: a clinicopathological analysis of 316 cases
Yan DONG ; Fanqing MENG ; Huidi HU ; Lihua SHEN ; Qianqian ZHANG ; Xiaoyan JIN ; Jue ZOU
Chinese Journal of Pathology 2025;54(11):1172-1179
Objective:To investigate the clinicopathological features, pathological classification, and molecular characteristics of pulmonary hamartomas.Methods:A retrospective analysis was conducted on 316 cases of pulmonary hamartomas diagnosed at Nanjing Chest Hospital, Nanjing, China from January 2015 to June 2024. Next generation sequencing (NGS) was performed on 15 cases of this study. The clinical data, histopathological features, immunophenotypes, and molecular alterations were analyzed. Relevant literature was reviewed.Results:Among the 316 patients, there were 154 males and 162 females, with an average age of 56±10 years. Among the 316 cases, 310 were intrapulmonary hamartomas and 6 were intraluminal bronchial hamartomas. Microscopically, there were complex proliferative mesenchymal components and epithelial components, presenting various combinations and hamartomatous morphologies. These hamartomas were morphologically classified into mesenchymal-type hamartomas (cartilaginous, fibrous, smooth muscle, adipose tissue, and mixed types) and epithelial-mesenchymal mixed-type hamartomas (respiratory epithelial-mesenchymal mixed and mucosal gland-mesenchymal mixed types). The cartilaginous hamartomas accounted for 72.8% (230/316) of them, and the non-cartilaginous hamartoma accounted for 27.2% (86/316). Secondary changes such as calcification, ossification, collagenization, mucin degeneration, and cystic changes were commonly present. The immunophenotype was CK7 +/TTF1 + for respiratory epithelial cells, or TTF1 -/CK7 +/p40 + for interstitial cells. Interstitial cells might express desmin, SMA, S-100, caldesmon, etc, while CD34 +/CD10 +/ER + spindle-shaped interstitial cells were also commonly noted. Genetic variations were detected in 11 of the 15 cases that were subject to NGS, including HMGA2-related fusion genes, EP300 mutations, FLT1 mutations, JAK1 mutations, SETD2 and TAP2 mutations, and high-copy amplification of CDK4/PHF1/TSPAN31. The patients were followed up for 6 to 110 months without any known recurrence or metastasis. Conclusions:Pulmonary hamartomas mainly occur in the peripheral lung parenchyma, with the cartilaginous type being the most common. Their clinical pathological and molecular features of pulmonary hamartomas are characterized and the histological types are roughly ascertained in this study, with emphasis of the key points of diagnosis and differential diagnosis. Classification of pulmonary hamartomas is valuable for guiding future research. Pulmonary hamartomas overall have a good prognosis. However, those with cystic changes or intraluminal hamartomas in the bronchus may cause serious airway lesions and therefore require special attention.
4.Direct measurement assessment study of internal exposure dose of nuclear medicine staff expose to 131I
Lihua HUANG ; Pan LIU ; Tianlu ZOU ; Xiaohua MIAO ; Qishan ZHENG ; Senxin ZHENG ; Weiqi WEI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2025;43(10):777-780
Objective:To understand the current situation of occupational internal exposure dose research using direct measurement method in China, and to explore the occupational internal dose level of nuclear medicine staff.Methods:From 2023 to April 2024, search on platforms such as China National Knowledge Infrastructure and Chinese Medical Journal Network, to collect research literature on the internal exposure dose monitoring of nuclear medicine staff and discuss the internal exposure dose assessment method. According to the literature reported thyroid 131I activity level at the measuring time, the internal exposure dose level was estimated using the method recommended by GBZ 129-2016. Results:According to existing research reports on direct measurement using portable γ spectrometers, 6.1% of nuclear medicine workers may receive a 131I dose greater than 1 mSv/a. Conclusion:It is necessary to conduct periodic continuous monitoring for personnel whose internal exposure dose may greater than 1 mSv/a, and it is necessary to explore the feasibility of periodic internal exposure monitoring method carried out by hospitals.
5.Exploratory study of Yisui Yangxin moxibustion in prevention and treatment of prehypertension in perimenopausal women.
Aixin HE ; Lihua ZHAO ; Zhuocheng ZOU ; Yu HUANG ; Dingjian HUANG
Chinese Acupuncture & Moxibustion 2025;45(8):1083-1091
OBJECTIVE:
To observe the effects of Yisui Yangxin moxibustion (moxibustion for benefiting the marrow and nourishing the heart) in sequential trial on blood pressure (BP), perimenopausal symptoms, cardiovascular function and heart rate variability (HRV) in prehypertension in perimenopausal women.
METHODS:
The eligible female patients of prehypertension of perimenopausal period were collected according to the inclusion criteria. Based on the requirements of open two-way qualitative response sequential trial, the sample size was not set in advance, and the matched pair design was used for random division into a moxibustion group and a health education group. The patients of the two groups were treated in pairs, and the analysis was immediately performed after the end of treatment for each pair, and the sequential trial analysis (STA) was diagrammed. The health education group provided the health education for prehypertension to the patients according to 2018 Revised Chinese Hypertension Prevention and Treatment Guidelines. In the moxibustion group, beside the health education for prehypertension, the non-suppurative moxibustion was delivered at Baihui (GV20), Guanyuan (CV4) and bilateral Neiguan (PC6) and Zusanli (ST36), once every two days, 3 treatments per week and for 4 consecutive weeks. Before treatment, in 2 and 4 weeks of treatment, as well as in 1-month follow-up after treatment, BP was measured in each group, separately. Before treatment and in 4 weeks of treatment, the observation was performed in the score of the modified Kupperman scale, and Pittsburgh sleep quality index (PSQI) score, the indexes of cardiovascular function (stroke volume [SV], cardiac output per minute [CO], stroke index [SI], cardiac index [CI], left ventricular effective pumping force [VPE], left ventricular energy efficiency [EWK], arterial compliance [AC], left ventricular ejection resistance [VER], blood viscosity (N), and microcirculation half update rate [MHR]), and heart rate variability (HRV) indexes (low frequency [LF], high frequency [HF], LF/HF, standard deviation of NN interval [SDNN], root mean square of successive differences in adjacent NN intervals [RMSSD]).
RESULTS:
Based on sequential test line, when the trial was performed to the 17th pair, and the test line touched the upper limit U, meaning the results of the moxibustion group was superior to the health education group, thus, the trial stopped immediately. In 2 and 4 weeks of treatment and in follow-up, the systolic blood pressure (SBP) and diastolic blood pressure (DBP) were lower when compared with those before treatment in the moxibustion group (P<0.05); SBP at each time point and DBP in 2 and 4 weeks of treatment in the moxibustion group were lower than those of the health education group (P<0.05). After treatment, the score of the modified Kupperman scale was reduced when compared with that before treatment in the moxibustion group (P<0.05). The score of the modified Kupperman scale and PSQI in the moxibustion group were lower than those of the health education group after treatment (P<0.05). After treatment, SV, CO, SI, CI and EWK were increased in comparison with the indexes before treatment in the moxibustion group (P<0.05), and VER and N were dropped (P<0.05). After treatment, SV, CO, SI, CI and EWK in the moxibustion group were higher than those of the health education group (P<0.05), and VER and N were lower (P<0.05). After treatment, in the moxibustion group, LF, HF, SDNN and RMSSD were increased in comparison with those before treatment (P<0.05), and LF/HF was declined (P<0.05). In the health education group, after treatment, LF, HF, SDNN and RMSSD decreased when compared with those before treatment (P<0.05). After treatment, LF, HF, SDNN and RMSSD in the moxibustion group were higher than those of the health education group (P<0.05), and LF/HF was lower (P<0.05). The total effective rate was 94.1% (16/17) in the moxibustion group, higher than that (41.2%, 7/17) in the health education group (P<0.05).
CONCLUSION
Moxibustion can reduce SBP and DBP in prehypertension of perimenopausal women, alleviate perimenopausal symptoms, improve cardiac function and adjust the overall balance of cardiac autonomic nerves.
Humans
;
Female
;
Middle Aged
;
Moxibustion
;
Perimenopause
;
Blood Pressure
;
Prehypertension/prevention & control*
;
Heart Rate
;
Acupuncture Points
;
Adult
6.Direct measurement assessment study of internal exposure dose of nuclear medicine staff expose to 131I
Lihua HUANG ; Pan LIU ; Tianlu ZOU ; Xiaohua MIAO ; Qishan ZHENG ; Senxin ZHENG ; Weiqi WEI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2025;43(10):777-780
Objective:To understand the current situation of occupational internal exposure dose research using direct measurement method in China, and to explore the occupational internal dose level of nuclear medicine staff.Methods:From 2023 to April 2024, search on platforms such as China National Knowledge Infrastructure and Chinese Medical Journal Network, to collect research literature on the internal exposure dose monitoring of nuclear medicine staff and discuss the internal exposure dose assessment method. According to the literature reported thyroid 131I activity level at the measuring time, the internal exposure dose level was estimated using the method recommended by GBZ 129-2016. Results:According to existing research reports on direct measurement using portable γ spectrometers, 6.1% of nuclear medicine workers may receive a 131I dose greater than 1 mSv/a. Conclusion:It is necessary to conduct periodic continuous monitoring for personnel whose internal exposure dose may greater than 1 mSv/a, and it is necessary to explore the feasibility of periodic internal exposure monitoring method carried out by hospitals.
7.Effects of cough enhancement techniques on weaning in adult patients under mechanical ventilation:a systematic review
Qian CHEN ; Junjun ZOU ; Chunyang XU ; Lihua GU ; Xiaojuan JIANG ; Mengqian GU
Modern Clinical Nursing 2025;24(4):81-91
Objective To systematically evaluate the impact of cough enhancement techniques on weaning outcomes in adult patients under mechanical ventilation(MV).Methods Databases from the inception to 15th November,2024 were searched for randomised controlled trials(RCTs)on cough enhancement technique in MV patients.The searched databases were CNKI,Wanfang Data base,SinoMed,Vip,PubMed,Web of Science,Cochrane Library,Embase,and CINAHL.Literature and evidence quality were evaluated using Cochrane's evaluation tool for intervention research and the GRADE system,and data were extracted and analysed via Meta-analysis with Review Manager 5.3.Results A total of 27 studies were included,involving 2,322 patients(1,169 in the trial group and 1,153 in the control group).Compared with the control group,the trial group had shorter time of mechanical ventilation[MD=-1.40,95%CI(-1.83,-0.97),P<0.001],lower weaning failure rate[OR=0.48,95%CI(0.27,0.85),P=0.01],higher oxygenation index[MD=35.48,95%CI(21.53,49.43),P<0.001],lower pulmonary atelectasis incidence[OR=0.30,95%CI(0.18,0.49),P<0.001],greater sputum volume[MD=1.88,95%CI(0.76,3.01),P=0.001],shorter ICU stay[MD=-1.90,95%CI(-2.95,-0.85),P<0.001]and lower serum procalcitonin(PCT)[MD=-0.89,95%CI(-1.64,-0.14),P=0.02],all with statistically significant differences.However,there was no significant difference in the ventilator-associated pneumonia(VAP)between the two groups[OR=0.54,95%CI(0.21,1.35),P=0.19].Conclusion Cough enhancement techniques promote sputum discharge,improve infection and oxygenation,shorten mechanical ventilation and ICU stay,reduce weaning failure and atelectasis rates in MV patients.Further studies are required to reduce the incidence of VAP.
8.Pulmonary hamartoma: a clinicopathological analysis of 316 cases
Yan DONG ; Fanqing MENG ; Huidi HU ; Lihua SHEN ; Qianqian ZHANG ; Xiaoyan JIN ; Jue ZOU
Chinese Journal of Pathology 2025;54(11):1172-1179
Objective:To investigate the clinicopathological features, pathological classification, and molecular characteristics of pulmonary hamartomas.Methods:A retrospective analysis was conducted on 316 cases of pulmonary hamartomas diagnosed at Nanjing Chest Hospital, Nanjing, China from January 2015 to June 2024. Next generation sequencing (NGS) was performed on 15 cases of this study. The clinical data, histopathological features, immunophenotypes, and molecular alterations were analyzed. Relevant literature was reviewed.Results:Among the 316 patients, there were 154 males and 162 females, with an average age of 56±10 years. Among the 316 cases, 310 were intrapulmonary hamartomas and 6 were intraluminal bronchial hamartomas. Microscopically, there were complex proliferative mesenchymal components and epithelial components, presenting various combinations and hamartomatous morphologies. These hamartomas were morphologically classified into mesenchymal-type hamartomas (cartilaginous, fibrous, smooth muscle, adipose tissue, and mixed types) and epithelial-mesenchymal mixed-type hamartomas (respiratory epithelial-mesenchymal mixed and mucosal gland-mesenchymal mixed types). The cartilaginous hamartomas accounted for 72.8% (230/316) of them, and the non-cartilaginous hamartoma accounted for 27.2% (86/316). Secondary changes such as calcification, ossification, collagenization, mucin degeneration, and cystic changes were commonly present. The immunophenotype was CK7 +/TTF1 + for respiratory epithelial cells, or TTF1 -/CK7 +/p40 + for interstitial cells. Interstitial cells might express desmin, SMA, S-100, caldesmon, etc, while CD34 +/CD10 +/ER + spindle-shaped interstitial cells were also commonly noted. Genetic variations were detected in 11 of the 15 cases that were subject to NGS, including HMGA2-related fusion genes, EP300 mutations, FLT1 mutations, JAK1 mutations, SETD2 and TAP2 mutations, and high-copy amplification of CDK4/PHF1/TSPAN31. The patients were followed up for 6 to 110 months without any known recurrence or metastasis. Conclusions:Pulmonary hamartomas mainly occur in the peripheral lung parenchyma, with the cartilaginous type being the most common. Their clinical pathological and molecular features of pulmonary hamartomas are characterized and the histological types are roughly ascertained in this study, with emphasis of the key points of diagnosis and differential diagnosis. Classification of pulmonary hamartomas is valuable for guiding future research. Pulmonary hamartomas overall have a good prognosis. However, those with cystic changes or intraluminal hamartomas in the bronchus may cause serious airway lesions and therefore require special attention.
9.Study on extraction technology of Sophora flavescens-Phellodendron chinense drug pair
Yongmei GUAN ; Ying TAO ; Chengyu ZOU ; Zhenzhong ZANG ; Lihua CHEN ; Lili LIU ; Limei CHEN ; Weifeng ZHU
China Pharmacy 2024;35(7):793-800
OBJECTIVE To study the extraction technology of Sophora flavescens-Phellodendron chinense drug pair and provide a reference for the development of new drugs for the treatment of anorectal diseases. METHODS Using the contents of total alkaloids of S. flavescens (matrine+oxymatrine), berberine hydrochloride and total flavonoid, and extract yield as evaluation indicators, analytic hierarchy process-entropy weight method was used to calculate the weight coefficient of each indicator, and was combined with Box-Behnken design-response surface method to study the extraction technology of S. flavescens-P. chinense drug pair and verify it. RESULTS The optimal extraction technology of S. flavescens-P. chinense drug pair was immersed in 12-fold amount of 58% ethanol for 30 minutes and extracted twice, each time for 120 minutes. The relative error between the verification experimental results and the predicted value was 1.88%. CONCLUSIONS The obtained extraction technology is stable and feasible and can provide reference for the application of S. flavescens-P. chinense drug pair and development of new drugs.
10.Visual analysis of hot topics in concussion field by finite element method:Improvements in brain injury models,test methods and protective devices
Weidong LUO ; Lihua ZOU ; Da HUANG
Chinese Journal of Tissue Engineering Research 2024;28(34):5487-5493
BACKGROUND:Concussions caused by contact sports or traffic accidents are far more serious and common than people think,and have attracted widespread attention and high attention from the media,medical and sports circles in recent years. OBJECTIVE:To visualize the hot spots and trends in the field of concussion by using the method of bibliometrics,so as to provide some reference for the research in this field in China. METHODS:Based on the core collection database of Web of Science,the keyword retrieval strategy was(TS=(Concussion))AND TS=(Finite element).CiteSpace 6.2.R4 visualization tool was used to visually analyze the author,country,institution,keywords,cited documents,etc. RESULTS AND CONCLUSION:A total of 215 articles were included,with a general upward trend in the number of publications and citations.The distribution of disciplines involves biomedical engineering,biophysics,sports science,clinical neurology,neuroscience and other disciplines,showing a trend of interdisciplinary integration.The author with the most publications is Gilchrist M from University College Dublin,Ireland,the institution with the most publications is the University of Ottawa,and the country with the most publications is the United States.Key word analysis shows that the focus of research is on the establishment of brain injury models to simulate and predict concussion injuries;analysis of concussion injury mechanism;optimal design of protective equipment and devices.Through literature co-citation analysis,it is found that the prediction and evaluation of brain injury is the knowledge base and research hotspot in this field.The research hotspot of finite element application in the field of concussion injury mainly focuses on the prediction of head injury,combined with the exploration of brain injury mechanism and the design and improvement of protective equipment.With the progress of artificial intelligence and materials science,future research hotspots in the field of concussion injury will focus on the improvement of brain injury models,test methods and protective equipment.

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