1.Construction of glioma microfluidic chip model and its application research on evaluation the medicinal efficacy of the Chinese medicine Scutellaria barbata
Piaoxue YOU ; Lan CHEN ; Yiwei SHI ; Hui WANG ; Liang CHAO ; Zhanying HONG
Journal of Pharmaceutical Practice and Service 2025;43(2):59-66
Objective To construct a glioma microfluidic chip model to simulate tumor microenvironment for evaluating the medicinal efficacy of anti-glioma traditional Chinese medicines. Methods Glioblastoma cells U251 were seeded into microfluidic chips with different culture modes, and the cell viability and tumour microenvironment within the constructed model were characterized. Fluorescence staining was used to evaluate the effects of the positive drugs temozolomide (TMZ) and docetaxel (DOC) on the cell activity and apoptosis within the model, which was applied to evaluate the medicinal efficacy of the extracts of the herb Scutellaria barbata on gliomas. Results The cells in the constructed U251 microfluidic chip model displayed high viability and were able to mimic the hypoxic microenvironment of tumor to a certain extent. The viability of the U251 cells in the microfluidic chips decreased with the increasing of the concentration of the positive drug, and the viability of the 3D cultured U251 cells was higher than that in the 2D condition (P<0.05). The intracellular mitochondrial membrane potential decreased with the increasing of the concentration of the positive drug. And the 2 mg/ml Scutellaria barbata extract killed U251 cells to a certain extent and reduced the mitochondrial membrane potential of the cells in the model. Conclusion This study successfully constructed a microfluidic chip model of glioma that could effectively simulate the tumor microenvironment and rapidly evaluate the anti-tumor medicinal efficacy, which provided a new strategy for the medicinal efficacy evaluation and active components screening of anti-glioma traditional Chinese medicines.
2.Factors influencing carbapenem-resistant gram-negative bacillus infection in elderly patients in the intensive care unit of a general hospital in Yangpu District, Shanghai, 2019‒2023
Wen ZHU ; Qingfeng SHI ; Yi LIANG ; Junping YU ; Yunxia LI ; Chao WENG ; Renyi ZHU
Shanghai Journal of Preventive Medicine 2025;37(6):467-475
ObjectiveTo analyze the characteristics and influencing factors of elderly hospitalized patients with carbapenem-resistant gram-negative bacillus (CRO) infection in the intensive care unit (ICU) of a gradeⅡ level A general hospital in Yangpu District of Shanghai, and to provide scientific basis for the prevention and control of hospital-acquired CRO infection in such hospitals. MethodsThe clinical data of elderly ICU patients (age ≥60 years) from January 2019 to December 2023 were retrospectively collected. A total of 122 cases with hospital-acquired CRO infection were used as the case group, and a total of 68 cases with carbapenem-sensitive gram-negative (CSO) infection were used as the control group. The clinical characteristics of the two groups were analyzed, and univariate analysis and logistic regression analysis were performed for screening for possible influencing factors on hospital-acquired CRO infection. ResultsThe main pathogens of CRO infection were carbapenem-resistant Acinetobacter baumannii (CRAB) (53 cases, 43.44%) and carbapenem-resistant Klebsiella pneumoniae (CRKP) (46 cases, 37.70%), and 17 patients (13.93%) had more than two types of CRO infection. Among the CRO infection, the main sites were lower respiratory tract infection (58 cases, 47.54%), ventilator-associated pneumonia (21 cases, 17.21%), and catheter-associated urinary tract infections (16 cases, 13.11%). The incidence rate of poor prognosis was higher in the CRO infection group (54.10%) than that in the CSO infection group (36.76%) (P=0.021). The results of univariate analysis showed that male, history of hospitalization within three months, chronic respiratory disease, hypoproteinemia, anemia, and history of invasive procedures prior to infection, including indwelling central venous catheter, invasive mechanical ventilation, urinary catheter, gastric tube placement and parenteral nutrition, in addition, heparin anticoagulation, the use of broad-spectrum penicillin, third-generation cephalosporins, fluoroquinolones, carbapenems, carbapenems combined with fluoroquinolones, carbapenems combined with glycopeptides, use of ≥3 antibiotics and long time of antibiotic use prior to infection were all associated with the CRO infection (P<0.05). The results of logistic regression analysis showed that use of carbapenems (OR=7.739, 95%CI: 2.226‒26.911), ≥3 types of antibiotics (OR=6.307, 95%CI: 1.674‒23.754), invasive mechanical ventilation (OR=4.082, 95%CI: 1.795‒9.281), urinary catheter (OR=3.554, 95%CI: 1.074‒11.758), and comorbid hypoproteinemia (OR=4.741, 95%CI: 2.039‒11.022) and diabetes (OR=3.245, 95%CI: 1.344‒7.839) were positively correlated with the risk of CRO infection. ConclusionConcurrent use of carbapenems with multiple other antibiotics, as well as the use of invasive mechanical ventilation, urinary catheter, and comorbid hypoproteinemia and diabetes, may be associated with an increased influencing of CRO infection. More attention should be paid to the prevention and control of infection in elderly patients with the above-mentioned risk factors, and active screening of drug-resistant bacteria should be strengthened. Besides, the rational use of broad-spectrum antibiotics such as carbapenems, avoiding unnecessary invasive operations, and paying attention to patient nutrition and blood glucose control all can reduce the incidence of CRO infection and help to improve clinical outcomes.
3.Risk factors and mortality for carbapenem-resistant Acinetobacter baumannii bloodstream infection in elderly patients:a 10-year retrospective study
Ye XUE ; Chao-Shi ZOU ; Tai-Jie LI ; Mei-Xiang QIN ; Chan LIANG ; Kang-Hai LIU ; Dan-Ping QIU
Chinese Journal of Infection Control 2024;23(2):155-161
Objective To assess the risk factors for carbapenem-resistant Acinetobacter baumannii(CRAB)bloodstream infection(BSI)and 28-day short-term mortality in elderly patients,and provide reference for the pre-vention and treatment of CRAB BSI.Methods Clinical data of patients aged ≥60 years and diagnosed with AB BSI in a hospital in Yulin City from January 2013 to December 2022 were retrospectively analyzed,including demogra-phic and microbiological characteristics,as well as clinical outcomes of the patients.Variables which were significant in univariate analysis were selected for multivariate analysis using binary logistic regression model and Cox propor-tional hazards model.Independent risk factors for infection were further determined,and survival analysis was per-formed using Kaplan-Meier curve.Results A total of 150 patients were included in the study,out of which 16 pa-tients(10.7%)had CRAB BSI and 134 had carbapenem-sensitive AB(CSAB)BSI.The 28-day short-term mortali-ty of AB BSI in elderly patients was 15.3%(23/150,95%CI:9.6%-21.1%),and the short-term mortality of CRAB BSI was higher than that of CSAB([56.3%,9/16]vs[10.4%,14/134]).Deep venous catheterization(OR:15.598,95%CI:1.831-132.910)and combined infections of other sites(OR:15.449,95%CI:1.497-159.489)were related to CRAB BSI in elderly patients.The independent risk factors for 28-day mortality in elderly patients with AB BSI were hemodialysis(OR:11.856,95%CI:2.924-48.076),intensive care unit admission(OR:9.387,95%CI:1.941-45.385),and pulmonary infection being suspected source of bacteremia(OR:7.019,95%CI:1.345-36.635).Conclusion The occurrence of CRAB BSI in elderly patients is related to the combined infection of other sites and deep vein catheterization.Hemodialysis,admission to ICU,and pulmonary infection being suspected source of bacteremia are independent risk factors for the prognosis of AB BSI in elderly patients.
4.Development of a High-throughput Sequencing Platform for Detection of Viral Encephalitis Pathogens Based on Amplicon Sequencing
Li Ya ZHANG ; Zhe Wen SU ; Chen Rui WANG ; Yan LI ; Feng Jun ZHANG ; Hui Sheng LIU ; He Dan HU ; Xiao Chong XU ; Yu Jia YIN ; Kai Qi YIN ; Ying HE ; Fan LI ; Hong Shi FU ; Kai NIE ; Dong Guo LIANG ; Yong TAO ; Tao Song XU ; Feng Chao MA ; Yu Huan WANG
Biomedical and Environmental Sciences 2024;37(3):294-302
Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.
5.Expert consensus on the biosafety recommendation for arthropods of medical importance in field and laboratory
HE Changhua ; LUO Huanle ; YIN Feifei ; HAN Qian ; LIANG Lei ; SHI Yongxia ; YU Xuedong ; SUN Yi ; LIU Qiyong ; WANG Huanyu ; WANG Rong ; SHAN Chao ; DENG Fei ; YUAN Zhiming ; XIA Han
China Tropical Medicine 2024;24(2):119-
The emerging and re-emerging arthropod-borne infectious diseases pose a serious threat to global public health security. Field and laboratory studies of arthropods of medical importance are essential and critical for the prevention and control of arthropod-borne infectious diseases. Various institutions or universities in China have been conducting research in the field or laboratory study of arthropods of medical importance, but up to 2023, it is still lacking detailed biosafety guidelines or recommendations that can guide the related work for arthropods of medical importance. In order to proactively address potential biosafety issues in the field or laboratory activities related to arthropods of medical importance, improve the standardization of arthropod biosafety classification, operations, and protection, and ensure the safety of practitioners, an expert consensus on the biosafety recommendation of arthropods of medical importance in field and laboratory has been developed, aiming to guide the future work of arthropods and ensure the national biosafety and biosecurity of China.
6.A retrospective study of occlusal reconstruction in patients with old jaw fractures and dentition defects
Ming-Chao DING ; Bo-Ya JING ; Jin SHI ; Liu YANG ; Xiang-Dong LIU ; Jing-Fu WANG ; Shuang QU ; Jia-Wu LIANG ; Zi-Hao TANG ; Jin-Long ZHAO ; Lei TIAN
Chinese Journal of Traumatology 2024;27(5):272-278
Purpose::This study evaluated the methods and clinical effects of multidisciplinary collaborative treatment for occlusal reconstruction in patients with old jaw fractures and dentition defects.Methods::Patients with old jaw fractures and dentition defects who underwent occlusal reconstruction at the Third Affiliated Hospital of Air Force Military Medical University from January 2018 to December 2022 were enrolled. Clinical treatment was classified into 3 phases. In phase I, techniques such as orthognathic surgery, microsurgery, and distraction osteogenesis were employed to reconstruct the correct 3-dimensional (3D) jaw position relationship. In phase II, bone augmentation and soft tissue management techniques were utilized to address insufficient alveolar bone mass and poor gingival soft tissue conditions. In phase III, implant-supported overdentures or fixed dentures were used for occlusal reconstruction. A summary of treatment methods, clinical efficacy evaluation, comparative analysis of imageological examinations, and satisfaction questionnaire survey were utilized to evaluate the therapeutic efficacy in patients with traumatic old jaw fractures and dentition defects. All data are summarized using the arithmetic mean ± standard deviation and compared using independent sample t-tests. Results::In 15 patients with old jaw fractures and dentition defects (an average age of 32 years, ranging from 18 to 53 years), there were 7 cases of malocclusion of single maxillary fracture, 6 of malocclusion of single mandible fracture, and 2 of malocclusion of both maxillary and mandible fractures. There were 5 patients with single maxillary dentition defects, 2 with single mandibular dentition defects, and 8 with both maxillary and mandibular dentition defects. To reconstruct the correct 3D jaw positional relationship, 5 patients underwent Le Fort I osteotomy of the maxilla, 3 underwent bilateral sagittal split ramus osteotomy of the mandible, 4 underwent open reduction and internal fixation for old jaw fractures, 3 underwent temporomandibular joint surgery, and 4 underwent distraction osteogenesis. All patients underwent jawbone augmentation, of whom 4 patients underwent a free composite vascularized bone flap (26.66%) and the remaining patients underwent local alveolar bone augmentation. Free gingival graft and connective tissue graft were the main methods for soft tissue augmentation (73.33%). The 15 patients received 81 implants, of whom 11 patients received implant-supported fixed dentures and 4 received implant-supported removable dentures. The survival rate of all implants was 93.82%. The final imageological examination of 15 patients confirmed that the malocclusion was corrected, and the clinical treatment ultimately achieved occlusal function reconstruction. The patient satisfaction questionnaire survey showed that they were satisfied with the efficacy, phonetics, aesthetics, and comfort after treatment.Conclusion::Occlusal reconstruction of old jaw fractures and dentition defects requires a phased sequential comprehensive treatment, consisting of 3D spatial jaw correction, alveolar bone augmentation and soft tissue augmentation, and implant-supported occlusal reconstruction, achieving satisfactory clinical therapeutic efficacy.
7.Analysis of the characteristics of infectious pathogens in burn patients with sepsis based on metagenomic next-generation sequencing technology
Jijing SHI ; Liang ZHAO ; Xiaoliang LI ; Qun ZHANG ; Chengde XIA ; Chao MA
Chinese Journal of Burns 2024;40(10):940-947
Objective:To analyze the characteristics of infectious pathogens in burn patients with sepsis based on metagenomic next-generation sequencing (mNGS) technology.Methods:This study was a retrospective observational study. From July 2021 to December 2023, 109 burn patients with sepsis who met the inclusion criteria were admitted to the Department of Burns of the First People's Hospital of Zhengzhou, including 68 males aged 57 to 92 years and 41 females aged 48 to 83 years. Blood, bronchoalveolar lavage fluid, cerebrospinal fluid, sputum, or other fluid specimens were collected from the patients during their hospital stay for microbiological culture (86 patients) and mNGS technology detection (109 patients). The types of specimens and pathogens detected by mNGS technology were counted. Patients were divided into intensive care unit (ICU) group (78 cases) who were admitted to the ICU and non-ICU group (31 cases) who were not admitted to the ICU, and the pathogens for infection in the two groups of patients were analyzed. In addition, the detection of pathogens in the specimens of 86 patients who underwent both mNGS technology detection and microbiological culture detection was analyzed.Results:Among the 109 specimens detected by mNGS technology, there were 42 blood specimens, 17 bronchoalveolar lavage fluid specimens, 4 sputum specimens, 6 cerebrospinal fluid specimens, 16 pus specimens, and 24 tissue fluid specimens; a total of 39 pathogens were detected, including 13 bacteria, 12 fungi, 10 viruses, 2 parasites, and 2 mycoplasmas. The overall positive rate of pathogen detection was 88.99% (97/109). Ranked by the detection rate, the top three Gram-negative bacteria were Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas spp, the top three Gram-positive bacteria were Streptococcus pneumoniae, Staphylococcus aureus, and Enterococcus faecalis; the top three viruses were human herpesvirus, cytomegalovirus, and circovirus; the top three fungi were Aspergillus fumigatus, Candida albicans, and Aspergillus flavus. Twenty-seven patients were infected with one pathogen, 45 patients with two pathogens, and 25 patients with three or more pathogens. Compared with those in non-ICU group, the proportions of Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas spp, Streptococcus pneumoniae, Aspergillus fumigatus, and cytomegalovirus detected in the patients in ICU group were significantly higher (with χ2 values of 8.62, 7.93, 3.93, 5.48, 4.28, and 5.58, respectively, P<0.05). In the pathogens detected by mNGS technology and microbiological culture method, the most common bacteria were Klebsiellapneumoniaeand Acinetobacter baumannii, and the most common fungi were strains of Aspergillus and Candida. There were 19 pathogens those could only be detected by mNGS technology, such as Lichtheimia ramosa, Pneumocystis jirovecii, Mycobacterium tuberculosis, viruses, etc.; there were no pathogens detected by microbiological culture method that couldn't be detected by mNGS technology. Compared with those detected by microbiological culture method, the overall positive rate, bacterial positive rate, and fungal positive rate detected by mNGS technology were significantly increased (with χ2 values of 45.52, 5.88, and 4.94, respectively, P<0.05). The 27.91% (24/86) of patients were detected positive by both methods, and 72.09% (62/86) of the patients were detected positive by mNGS technology but negative by microbiological culture method. The consistency test of the results obtained by the two detection methods showed that the difference was not statistically significant ( κ=0.02, P>0.05). Conclusions:The positive rate of pathogen detection in specimens using mNGS technology is higher than that detected by using conventional microbiological culture method, and it can detect pathogens those cannot be detected by the latter, such as Lichtheimia ramosa, Pneumocystis jirovidii, Mycobacterium tuberculosis, viruses, etc. Detection using mNGS technology can help clarify the types of infectious pathogens in burns patients with sepsis, and provide basis and guidance for clinical medication.
8.Overexpression of Hsp70 Promoted the Expression of Glycolysis-related Genes in C2C12 Cells
Lei QIN ; Ke XU ; Chun-Guang ZHANG ; Han CHU ; Shi-Fan DENG ; Jian-Bin ZHANG ; Hua YANG ; Liang HONG ; Gui-Feng ZHANG ; Chao SUN ; Lei PU
Chinese Journal of Biochemistry and Molecular Biology 2024;40(10):1417-1425
The aim of this study was to investigate the impact of overexpressing 70-kD heat shock pro-teins(Hsp70)on glycolysis in C2C12 cells during myogenesis and adipogenesis.Using C2C12 cells as the research material,adenovirus was used to overexpress the Hsp70 gene,and changes in the expression of glycolytic genes were detected using fluorescence quantitative PCR and Western blotting techniques.The study indicated that during C2C12 cell myogenic differentiation,the expression trend of the Hsp70 gene was consistent with that of Gsk3β,Pkm,Prkag3,Pfkm,and Hk-2 genes,suggesting a relationship between Hsp70 and the glycolytic pathway during myogenic differentiation.Overexpression of Hsp70 in the later stages of myogenic differentiation significantly upregulated the expression of Gsk3β,Pkm,Prk-ag3,and Pfkm genes(P<0.05),with no significant impact on Hk-2 gene expression(P>0.05).Dur-ing C2C12 cell adipogenic induction,the expression trend of the Hsp70 gene was similar to that of Gsk3β,Pkm,Prkag3,Pfkm,and Hk-2 genes,indicating a relationship between Hsp70 and the glycolytic path-way during adipogenic induction.Following Hsp70 overexpression,in the later stages of adipogenic in-duction,the number of lipid droplets was significantly higher compared to the control group,with a sig-nificant upregulation of Gsk3β,Pkm,Prkag3,and Pfkm gene expression(P<0.05),while Hk-2 gene expression was not significantly affected(P>0.05).In conclusion,Hsp70 in C2C12 cells in myogenic and adipogenic states promoted the breakdown of glycogen into 6-phospho-glucose,thereby enhancing the glycolytic pathway,providing insights into the functional role of the Hsp70 gene in glycolysis in C2C12 cells.
9.Study of pre-hospital temporary VV-ECMO for the treatment of high-altitude severe blast lung injury based on animal models
Zheng-Bin WU ; Shi-Feng SHAO ; Liang-Chao ZHANG ; Zhao-Xia DUAN ; Yao-Li WANG ; Zong-An LIANG ; Jian-Min WANG
Journal of Regional Anatomy and Operative Surgery 2024;33(5):373-378
Objectives To explore the feasibility of temporary veno-venous extracorporeal membrane oxygenation(VV-ECMO)technology for early on-site treatment,through establishing an animal model of severe blast lung injury in goats by free-field chemical explosion experiments in high-altitude regions.Methods A total of 16 adult goats were selected,and divided into the control group and the treatment group according to the random number table method,with 8 goats in each group.A model of severe blast lung injury was established at an altitude of 4 600 meters above sea level,then the goats in the control group were given respiratory support and the goats in the treatment group were given temporary VV-ECMO treatment.The survival status of the goats 15 minutes after injury was recorded,the vital signs[including body temperature,respiration rate,heart rate,and mean arterial pressure(MAP)]and arterial blood gas analysis indicators[including pH,arterial partial pressure of oxygen(PO2),arterial partial pressure of carbon dioxide(PCO2),oxygen saturation(SaO2),lactate(LAC),calcium(Ca2+),hematocrit(HCT),and hemoglobin(Hb)]before injury and 1 hour,2 hours,3 hours after injury were compared in the two groups.The post-mortem examination was performed on all dead goats and sacrificed goats after treatment,the severity of lung injury was assessed by organ injury scaling(OIS),and the lung injury score was evaluated by abbreviated injury scale(AIS).The wet-to-dry weight ratio(W/D)and lung coefficient were calculated.Results Within 15 minutes after the explosion,4 goats in the control group died and 4 goats survived;and 5 goats in the treatment group died and 3 goats survived.There was no statistically significant difference in the body temperature,respiration rate,heart rate,or MAP before and after injury between the two groups(P>0.05).The PaO2 and SaO2 1 hour,2 hours,and 3 hours after injury in the treatment group were superior than those in the control group(P<0.05),the Ca2+ 2 hours after injury was significantly higher than that in the control group(P<0.05),and there was no statistically significant difference in the pH,PCO2,LAC,HCT or Hb at different time points after injury between the two groups(P>0.05).There was no statistically significant difference in the OIS,AIS or lung coefficient between the two groups(P>0.05),but the W/D of the lung tissue in the control group was lower than that in the treatment group(P<0.05).Conclusion We have established a novel,feasible,and stable treatment effect temporary VV-ECMO animal treatment strategy for the first time in the high-altitude regions,which can provide animal experiment evidence for the early on-site VV-ECMO treatment of severe blast lung injury in high-altitude regions.
10.The prevention and management of stent under-expansion
Hao JIANG ; Min SUN ; Chao LIANG ; Tian TIAN ; Shi-Jie FANG ; Qiang TANG
Chinese Journal of Interventional Cardiology 2024;32(10):596-600
Stent under-expansion is a relatively common situation during coronary intervention procedure,which can lead to serious consequences such as stent restenosis,stent thrombosis and stent rupture.How to early identify and deal with stent under-expansion has achieved widespread attention in recent years.Intraluminal imaging guided percutaneous coronary intervention plays an important role in the prevention of stent under-expansion.Besides,recent studies have demonstrated the feasibility of shockwave balloon,excimer laser coronary atherectomy and rotational atherectomy in dealing with stent under-expansion.This paper reviews the pathophysiology,intraluminal imaging evaluation and management of stent under-expansion to provide a reference for coronary intervention.

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