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.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.
4.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.
5.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.
6.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
7.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.
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.Examining core symptoms and influencing factors of patients with gastric cancer undergoing chemotherapy:the role of Traditional Chinese Medicine constitutional traits
Yanling ZOU ; Yi LI ; Ziyan ZHANG ; Xun LI ; Lihua YANG ; Xiaoqing WANG ; Juan GAO ; Huiyun GUAN ; Peibei DUAN
Chinese Journal of Nursing 2024;59(18):2238-2243
Objective To investigate the incidence and severity of symptoms in patients with gastric cancer who received chemotherapy,we constructed a symptom network to explore core symptoms and bridge symptoms.Furthermore,the study explores the association between core symptoms and Traditional Chinese Medicine(TCM)constitutions.Methods Patients with gastric cancer who received chemotherapy in the medical oncology and surgical oncology department from March to August 2023 were selected for the study using a convenience sampling method.The MD Anderson Symptom Inventory Gastrointestinal Cancer was used for evaluating gastrointestinal symptoms and their severity among patients receiving chemotherapy for gastric cancer,as well as assessing the classification of TCM constitution among patients.The symptom network model was constructed using the R programming language,and the central index was analyzed to determine the core symptoms and bridge symptoms.Binary logistic regression analysis was employed to assess the association between different physical conditions and the occurrence of core symptoms.Results A total of 346 electronic questionnaires were collected,with 340 valid ones,and the effective recovery rate was 98.3%.The 3 most prevalent and severe symptoms among the 340 patients with gastric cancer were fatigue(85.59%),lack of appetite(82.35%),and taste alteration(81.18%).The centrality index results indicated that grief exhibited the highest intensity,medium,and compactness centrality values(rs=8.23,rb=2.00,rc=0.03),making it the core symptom of this condition.Sleep disorders,lack of appetite,drowsiness,and taste alteration were identified as bridging symptoms with bridge intensities of 0.74,0.76,0.99,and 0.94 respectively.The results of Spearman correlation analysis showed that there was a positive correlation between sadness and qi-deficiency constitution,phlegm-dampness constitution(P<0.05).The phlegm-dampness constitution was positively correlated with the taste alterations(P<0.05).Conclusion In patients with gastric cancer,fatigue emerges as the most prominent symptom,while sadness assumes the core symptom.Additionally,sleep disorder,lack of appetite,drowsiness,and taste alteration are bridge symptoms.According to the principles of TCM constitution,qi-deficiency and phlegm-dampness are constitutions associated with a higher risk of experiencing sadness,and phlegm-dampness is a constitution associated with a higher risk of taste changes.Nurses can integrate core symptoms and TCM constitutions characteristics to optimize the strategies for symptom intervention.
10.Antibiotic resistance and epidemiological characteristics of polymyxin-resistant Klebsiella pneumoniae
Lihua CHEN ; Mengqian DENG ; Jiali WANG ; Tianrui WU ; Shenghong ZHOU ; Ruyin YANG ; Di ZHANG ; Mingxiang ZOU
Journal of Central South University(Medical Sciences) 2024;49(5):737-747
Objective:The emergence of polymyxin-resistant Klebsiella pneumoniae(KPN)in clinical settings necessitates an analysis of its antibiotic resistance characteristics,epidemiological features,and risk factors for its development.This study aims to provide insights for the prevention and control of polymyxin-resistant KPN infections. Methods:Thirty clinical isolates of polymyxin-resistant KPN were collected from the Third Xiangya Hospital of Central South University.Their antibiotic resistance profiles were analyzed.The presence of carbapenemase KPC,OXA-48,VIM,IMP,and NDM was detected using colloidal gold immunochromatography.Hypervirulent KPN was initially screened using the string test.Biofilm formation capacity was assessed using crystal violet staining.Combination drug susceptibility tests(polymyxin B with meropenem,tigecycline,cefoperazone/sulbactam)were conducted using the checkerboard method.Polymyxin-related resistance genes were detected by PCR.Multi-locus sequence typing(MLST)was performed for genotyping and phylogenetic tree construction.The study also involved collecting data from carbapenem-resistant(CR)-KPN polymyxin-resistant strains(23 strains,experimental group)and CR-KPN polymyxin-sensitive strains(57 strains,control group)to analyze potential risk factors for polymyxin-resistant KPN infection through univariate analysis and multivariate Logistic regression.The induction of resistance by continuous exposure to polymyxin B and colistin E was also tested. Results:Among the 30 polymyxin-resistant KPN isolates,28 were CR-KPN,all producing KPC enzyme.Four isolates were positive in the string test.Most isolates showed strong biofilm formation capabilities.Combination therapy showed additive or synergistic effects.All isolates carried the pmrA and phoP genes,while no mcr-1 or mcr-2 genes were detected.MLST results indicated that ST11 was the predominant type.The phylogenetic tree suggested that polymyxin-resistant KPN had not caused a hospital outbreak in the institution.The use of two or more different classes of antibiotics and the use of polymyxin were identified as independent risk factors for the development of polymyxin-resistant strains.Continuous use of polymyxin induced drug resistance. Conclusion:Polymyxin-resistant KPN is resistant to nearly all commonly used antibiotics,making polymyxin-based combination therapy a viable option.No plasmid-mediated polymyxin-resistant KPN has been isolated in the hospital.Polymyxin can induce resistance in KPN,highlighting the need for rational antibiotic use in clinical settings to delay the emergence of resistance.

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