1.Esophageal squamous cell carcinoma exosomal miR-181b-5p promotes tumor associated macrophage polarization by inhibiting PTEN
Hui FAN ; Naikuo ZHAO ; Linlin CHEN ; Zhiguo LI ; Miao LIU ; Yuanying ZHANG ; Chaofeng ZHOU
Chinese Journal of Immunology 2024;40(7):1441-1446
Objective:To investigate the effect and mechanism of esophageal squamous cell carcinoma exosomal miR-181b-5p to the polarization of M2 macrophages.Methods:Extracted and identified exosomes from esophageal squamous cell carcinoma,and detected the expression of miR-181b-5p in esophageal squamous cell carcinoma cells and their exosomes by qRT-PCR.M0 type macro-phages were divided into PBS group,HEEC exo group,Eca-109 exo group,miR-NC exo group,miR-181b-5p exo group,miR-NC group,miR-181b-5p mimic group,si-NC group,si-PTEN group,miR-181b-5p exo+PTEN group.qRT-PCR was used to detect the expressions of CD163,CD206,iNOS and TNF-α in each group.The targeting relationship between miR-181b-5p and PTEN were veri-fied by double luciferase reporter gene experiment.Results:miR-181b-5p was significantly overexpressed in esophageal squamous cell carcinoma cells TE-13,TE-12,TE-10,Eca-109,KYSE30 and their exosomes(P<0.001).Compared with miR-NC group,the expression of CD163 and CD206 in cells were significantly upregulated in the miR-181b-5p mimic group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).The results of double luciferase reporter genes showed that PTEN was the target gene of miR-181b-5p.Compared with si-NC group,the expressions of CD163 and CD206 in cells were significantly upregu-lated in the si-PTEN group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).Compared with PBS group,the expressions of CD163 and CD206 in cells were significantly upregulated in the Eca-109 exo group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).Compared with miR-NC exo group,the expressions of CD163 and CD206 in cells were significantly upregulated in the miR-181b-5p exo group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).Compared with miR-181b-5p exo group,the expressions of CD163 and CD206 in cells were significantly downregulated in the miR-181b-5p exo+PTEN group,as well as the expressions of iNOS and TNF-α were significantly upregulated(P<0.001).Conclusion:Exosomal miR-181b-5p inhibits PTEN expressions to promote M2 macrophage polarization.
2.Short term efficacy on 3D printing assisted reconstruction of traumatic digit joint defects using rib and costoosteochondral autograft
Chaofeng XING ; Zhiyu HU ; Xiazhi LIU ; Tao YANG ; Jia CHEN ; Zirun XIAO ; Li SONG ; Beibei CHENG ; Yingjie XIONG ; Guangchao ZHANG ; Yongsheng HE ; Gaowei ZHANG
Chinese Journal of Microsurgery 2024;47(3):280-286
Objective:To explore the short-term efficacy on 3D printing assisted reconstruction of traumatic digit joint defects using rib and costo-osteochondral autograft.Methods:From August 2022 to July 2023, 7 patients with open digit joint defects had undergone emergency primary debridement and fracture fixation in the Department of Orthopaedics, 988th Hospital of the Joint Logistics Support Force of PLA. Patients with more phalangeal defects that could not be aligned were treated with antibiotic bone cement filling in the emergency surgery. In the second stage surgery, bone cement was removed and transfer of rib cartilage graft was performed to reconstruct the digit joint defect. According to a 1∶1 3D printed hand templates, rib cartilage grafts were crafted to the shape of digit joints, and then spliced together the digit joints and bone defects for fixation. Follow-up X-ray examinations were taken and assessment of the healing status of rib and fractures of phalangeal and metacarpophalangeal bones were carried out according to the Paley fracture healing score. At the outpatient follow-up, assessment of transferred joint movement and evaluation of upper limb function were conducted according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association. Visual Analogue Scale (VAS) pain scores were evaluated from the affected digits and donor sites.Results:After reconstructive surgery, all 7 patients had primary healing of the wounds of hand. One patient had fat liquefaction at the donor site, and the rest had primary donor site healing. One patient received further surgery for extensor tendon repair after rib cartilage grafting due to the digital extensor tendon injury. All 7 patients were included in postoperative follow-up for 6-11 months, with an average of 9 months. All patients had excellent fracture healing according to the Paley fracture healing score. At the final follow-up, the extension and flexion of the digit joints were found at 40°-80° (average 56.2°) for proximal interphalangeal joints (4 patients), and 10° in extension and 85° in flexion for metacarpophalangeal joint (1 patient). The range of motion of the thumb interphalangeal joint (2 patients) was 20°-35° (average 27.5°). Hand function was assessed according to Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association and it was found that 3 patients were in excellent, 3 in good and 1 in fair.Conclusion:This study focused on the treatment of traumatic digit joint defects by transfer of individually crafted rib cartilages in reconstruction of the defected digit joint. It significantly improves the appearance and function of the defected digit joints, especially suitable for the irregular defects of phalangeal bones.
3.Clinical effect of trapezoidal transparent corneal incision during phacoemulsi-fication
Chaofeng YUAN ; Fei CHEN ; Wenna GAO ; Chengxia MA ; Xiaohua ZHAO ; Ling ZHANG ; Fengyan ZHANG
Recent Advances in Ophthalmology 2024;44(8):619-622
Objective To explore the clinical effect of a trapezoidal transparent corneal incision during phacoemulsi-fication.Methods A total of 57 patients(68 eyes)undergoing phacoemulsification were selected and divided into a con-ventional incision group and a trapezoidal incision group using a random number table method.There were 28 patients(34 eyes)in the conventional incision group,including 15 eyes(males)and 19 eyes(females),with an age range of 41-82(65.0±10.1)years;and there were 29 patients(34 eyes)in the trapezoidal incision group,including 21 eyes(males)and 13 eyes(females),with an age range of 46-87(66.0±11.1)years.All patients underwent cataract phacoemulsification combined with intraocular lens(IOL)implantation.A 3.0 mm transparent corneal incision was made for patients in the conventional incision group,while an improved trapezoidal transparent corneal incision was made for patients in the trape-zoidal incision group.The uncorrected visual acuity,intraocular pressure,corneal astigmatism,and incidence of intraoper-ative and postoperative complications were compared between the two groups.Results The uncorrected visual acuity of patients in the trapezoidal incision group was better than that of patients in the conventional incision group at 1 week and 3 months after surgery,and the differences were statistically significant(both P<0.05).There was no statistically significant difference in intraocular pressure and corneal astigmatism between the two groups of patients at 1 week,1 month,and 3 months after surgery(all P>0.05).The number of eyes with anterior chamber collapse and the number of eyes requiring a watertight incision in the conventional incision group were greater than those in the trapezoidal incision group during sur-gery,and the differences were statistically significant(both P<0.05).The number of eyes with incision edema and the number of eyes with incision gap in the conventional incision group were greater than those in the trapezoidal incision group after surgery,and the differences were statistically significant(both P<0.05).In the conventional incision group,IOL was displaced in 18 eyes due to the shallow anterior chamber and then returned to normal after the formation of the anterior chamber through a watertight incision during surgery;during the formation,iris prolapse and incarceration occurred in 2 eyes,and IOL rotation or incarceration occurred in 3 eyes.In the trapezoidal incision group,3 eyes had a shallow anterior chamber after surgery,and a watertight incision was used to form the anterior chamber;there was no IOL incarceration or displacement or iris prolapse.Conclusion The improved trapezoidal transparent corneal incision can effectively prevent IOL displacement caused by anterior chamber collapse during cataract surgery while ensuring the stability of the anterior chamber.It can also reduce the related complications caused by the watertight incision using a flushing needle and restore patients'vision as early as possible.
4.The role and regulatory mechanism of RNA binding protein ZFP36in hypoxia/reoxygenation injury of cardiomyocytes
Guo LÜ ; Chaofeng SUN ; Hao ZHANG ; Hongjun LI ; Fang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(2):219-227
【Objective】 To explore the role of ZFP36 in cardiomyocyte injury and autophagy induced by hypoxia/reoxygenation (H/R) so as to clarify its molecular regulatory mechanism. 【Methods】 H9C2 rat cardiomyocytes were infected with ZFP36 overexpressing lentivirus (OE-ZFP36) or its negative control lentivirus (OE-ZFP36 NC) to construct stable cell lines, respectively. Transfection of ATG4D overexpression plasmid (OE-ATG4D) improved the expression of ATG4D. Hypoxia/reoxygenation (H/R) induced myocardial cell injury. H9C2 cells were mainly divided into control group, H/R group, OE-ZFP36 NC+H/R group, OE-ZFP36+H/R group, OE-ATG4D NC+H/R group, OE-ATG4D+H/R group, OE-ZFP36+OE-ATG4D NC+H/R group, and OE-ZFP36+OE-ATG4D+H/R group. The protein expressions of ATG4D, Beclin1, LC3 and ZFP36 in H9C2 cells were detected by Western blotting. The mRNA levels of ZFP36 and ATG4D in H9C2 cells were detected by Real-time fluorescence quantitative PCR (qPCR). The viability of H9C2 cells was detected by CCK-8 assay. The levels of interleukin (IL-6) and tumor necrosis factor (TNF-α) in H9C2 cells were detected by enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) in H9C2 cells were detected by DCFH-DA method. SOD detection kit was used to detect the SOD level in H9C2 cells. The apoptosis of H9C2 cells was detected by flow cytometry. LC3 autophagosomes in H9C2 cells were detected by cellular immunofluorescence. Dual-luciferase reporter gene assay was used to detect the binding of ZFP36 and ATG4D mRNA in H9C2 cells. 【Results】 Compared with control group, H/R group showed decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels and decreased SOD levels, increased cell apoptosis. Up-regulated ATG4D and Beclin1 protein expression, increased LC3Ⅱ/LC3Ⅰ ratio, as well as upregulated ZFP36 expression were found in H/R group (all P<0.05). Compared with OE-ZFP36 NC+H/R group, elevated cell viability, decreased IL-6 and TNF-α levels, decreased ROS levels and increased SOD levels, reduced cell apoptosis (P<0.05), and downregulated ATG4D and Beclin1 protein expression, decreased LC3Ⅱ/LC3Ⅰ ratio were shown in OE-ZFP36+H/R group (all P<0.05). Compared with infection with OE-ZFP36 NC lentivirus, infection with OE-ZFP36 lentivirus decreased the luciferase activity of ATG4D 3′-UTR reporter gene, decreased the stability of ATG4D mRNA, and downregulated the H/R-induced ATG4D mRNA expression (all P<0.05). Compared with OE-ATG4D NC+H/R group, OE-ATG4D+H/R group had upregulated ATG4D mRNA and protein expression, decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). Compared with OE-ZFP36+OE-ATG4D NC+H/R group, OE-ZFP36+OE-ATG4D+H/R group had decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). 【Conclusion】 The expression of ZFP36 is upregulated in H/R-induced cardiomyocyte injury. The overexpression of ZFP36 inhibits H/R-induced cardiomyocyte injury and autophagy by regulating ATG4D, thus resisting cardiomyocyte H/R injury. It proves that ZFP36 is an important regulatory molecule against MI/RI.
5.Prediction of the clinical supply of blood components in Xi′an City from 2023 to 2025
Linghao ZHANG ; Jin WANG ; Bei YAO ; Xiaoyue CHU ; Zhendong SUN ; Chaofeng MA
Chinese Journal of Preventive Medicine 2024;58(8):1213-1218
Objective:To construct a prediction model for the clinical supply of blood components in Xi′an City from 2023 to 2025.Methods:Based on the blood supply data of the Blood Management Information System of Shaanxi Provincial Blood Center from January 2013 to December 2022, a gray prediction model and an exponential curve fitting model were used to construct the prediction model, and the optimal prediction model was determined according to the error parameters of the relevant indicators of the model. The supply of blood components in Xi′an from 2023 to 2025 was predicted.Results:The fitting equations of the exponential curve fitting model to predict the supply of suspended red blood cells, platelets and cryoprecipitate in Xi′an were, x(1)( t+1)=1.16e 0.04t, x(1)( t+1)=1.04e 0.12t and x(1)( t+1)=1.01e 1.10t, respectively. The mean absolute errors (mean relative errors) of the exponential curve fitting model in predicting the supply of suspended red blood cells, platelets and cryoprecipitate in Xi′an were 10 488.7 (0.05%), 2 114.9 (0.08%) and 3 089.6 (0.07%), respectively, which were lower than those of the gray prediction model, about 10 488.7 (3.44%), 2 152.78 (8.20%) and 3 441.35 (7.92%), respectively. The exponential curve fitting model predicted that the clinical supply of blood components in Xi′an would increase year by year from 2023 to 2025, and the clinical supply of suspended red blood cells, platelets, and cryoprecipitate in Xi′an would increase to 409 467 U, 69 818 therapeutic volume and 94 724 U, respectively by 2025. Conclusion:The exponential curve fitting model can make a good prediction of the clinical supply of blood components in Xi′an City.
6.Prediction of the clinical supply of blood components in Xi′an City from 2023 to 2025
Linghao ZHANG ; Jin WANG ; Bei YAO ; Xiaoyue CHU ; Zhendong SUN ; Chaofeng MA
Chinese Journal of Preventive Medicine 2024;58(8):1213-1218
Objective:To construct a prediction model for the clinical supply of blood components in Xi′an City from 2023 to 2025.Methods:Based on the blood supply data of the Blood Management Information System of Shaanxi Provincial Blood Center from January 2013 to December 2022, a gray prediction model and an exponential curve fitting model were used to construct the prediction model, and the optimal prediction model was determined according to the error parameters of the relevant indicators of the model. The supply of blood components in Xi′an from 2023 to 2025 was predicted.Results:The fitting equations of the exponential curve fitting model to predict the supply of suspended red blood cells, platelets and cryoprecipitate in Xi′an were, x(1)( t+1)=1.16e 0.04t, x(1)( t+1)=1.04e 0.12t and x(1)( t+1)=1.01e 1.10t, respectively. The mean absolute errors (mean relative errors) of the exponential curve fitting model in predicting the supply of suspended red blood cells, platelets and cryoprecipitate in Xi′an were 10 488.7 (0.05%), 2 114.9 (0.08%) and 3 089.6 (0.07%), respectively, which were lower than those of the gray prediction model, about 10 488.7 (3.44%), 2 152.78 (8.20%) and 3 441.35 (7.92%), respectively. The exponential curve fitting model predicted that the clinical supply of blood components in Xi′an would increase year by year from 2023 to 2025, and the clinical supply of suspended red blood cells, platelets, and cryoprecipitate in Xi′an would increase to 409 467 U, 69 818 therapeutic volume and 94 724 U, respectively by 2025. Conclusion:The exponential curve fitting model can make a good prediction of the clinical supply of blood components in Xi′an City.
7.Research Progress on Dendrobii Officinalis Caulis as Medicinal and Edible Traditional Chinese Medicine
Yang HU ; Mian ZHAO ; Yuxuan QIU ; Debao YE ; Yangqing LIU ; Chaofeng ZHANG ; Haibo WANG ; Jian-Ming CHENG
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(1):94-108
Tiepishihu(Dendrobii officinalis Caulis)is a medicinal and food source herbal medicine with the effect of benefiting stomach and promoting fluid,nourishing Yin and clearing heat.It has rich chemical components and pharmacological activities,with anti-inflammatory,anti-bacterial,anti-oxidation,anti-tumor,immunomodulatory,blood press regulation,hypoglycemic effects.It is not only used as medicinal food and health care products,but also widely used in medicine,such as Shihu Yeguang Wan,Compound fresh dendrobium granules and other drugs,with high medicinal and economic value.This paper summarized the resource distribution,chemical composition,pharmacological activities,and medicine and food of Dendrobii officinalis Caulis,and analyzed its application status,laying a theoretical foundation for the sustainable development,medicinal and food homologous development and comprehensive utilization of Dendrobii officinalis Caulis.
8.Recombinant expression of Japanese encephalitis virus non-structural protein NS1 gene and its reaction with Flavivirus antigen and antibody
ZHANG Yijia ; YAO Xiaohui ; CAO Lei ; WANG Ruichen ; FU Shihong ; NIE Kai ; LI Fan ; YIN Qikai ; HE Ying ; WANG Huanyu ; XU Songtao ; MA Chaofeng ; LIANG Guodong
China Tropical Medicine 2023;23(12):1241-
Abstract: Objective To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.
9.Promotion effect of FGF23 on osteopenia in congenital scoliosis through FGFr3/TNAP/OPN pathway.
Hongqi ZHANG ; Gang XIANG ; Jiong LI ; Sihan HE ; Yunjia WANG ; Ang DENG ; Yuxiang WANG ; Chaofeng GUO
Chinese Medical Journal 2023;136(12):1468-1477
BACKGROUND:
Congenital scoliosis (CS) is a complex spinal malformation of unknown etiology with abnormal bone metabolism. Fibroblast growth factor 23 (FGF23), secreted by osteoblasts and osteocytes, can inhibit bone formation and mineralization. This research aims to investigate the relationship between CS and FGF23.
METHODS:
We collected peripheral blood from two pairs of identical twins for methylation sequencing of the target region. FGF23 mRNA levels in the peripheral blood of CS patients and age-matched controls were measured. Receiver operator characteristic (ROC) curve analyses were conducted to evaluate the specificity and sensitivity of FGF23. The expression levels of FGF23 and its downstream factors fibroblast growth factor receptor 3 (FGFr3)/tissue non-specific alkaline phosphatase (TNAP)/osteopontin (OPN) in primary osteoblasts from CS patients (CS-Ob) and controls (CT-Ob) were detected. In addition, the osteogenic abilities of FGF23-knockdown or FGF23-overexpressing Ob were examined.
RESULTS:
DNA methylation of the FGF23 gene in CS patients was decreased compared to that of their identical twins, accompanied by increased mRNA levels. CS patients had increased peripheral blood FGF23 mRNA levels and decreased computed tomography (CT) values compared with controls. The FGF23 mRNA levels were negatively correlated with the CT value of the spine, and ROCs of FGF23 mRNA levels showed high sensitivity and specificity for CS. Additionally, significantly increased levels of FGF23, FGFr3, OPN, impaired osteogenic mineralization and lower TNAP levels were observed in CS-Ob. Moreover, FGF23 overexpression in CT-Ob increased FGFr3 and OPN levels and decreased TNAP levels, while FGF23 knockdown induced downregulation of FGFr3 and OPN but upregulation of TNAP in CS-Ob. Mineralization of CS-Ob was rescued after FGF23 knockdown.
CONCLUSIONS
Our results suggested increased peripheral blood FGF23 levels, decreased bone mineral density in CS patients, and a good predictive ability of CS by peripheral blood FGF23 levels. FGF23 may contribute to osteopenia in CS patients through FGFr3/TNAP / OPN pathway.
Humans
;
Osteopontin/genetics*
;
Alkaline Phosphatase/metabolism*
;
Receptor, Fibroblast Growth Factor, Type 3/metabolism*
;
Scoliosis/genetics*
;
Osteoblasts/metabolism*
;
Calcinosis
;
RNA, Messenger/metabolism*
;
Bone Diseases, Metabolic/metabolism*
;
Fibroblast Growth Factors/genetics*
10.Impaired autophagy activity-induced abnormal differentiation of bone marrow stem cells is related to adolescent idiopathic scoliosis osteopenia.
Hongqi ZHANG ; Guanteng YANG ; Jiong LI ; Lige XIAO ; Chaofeng GUO ; Yuxiang WANG
Chinese Medical Journal 2023;136(17):2077-2085
BACKGROUND:
Osteopenia has been well documented in adolescent idiopathic scoliosis (AIS). Bone marrow stem cells (BMSCs) are a crucial regulator of bone homeostasis. Our previous study revealed a decreased osteogenic ability of BMSCs in AIS-related osteopenia, but the underlying mechanism of this phenomenon remains unclear.
METHODS:
A total of 22 AIS patients and 18 age-matched controls were recruited for this study. Anthropometry and bone mass were measured in all participants. Bone marrow blood was collected for BMSC isolation and culture. Osteogenic and adipogenic induction were performed to observe the differences in the differentiation of BMSCs between the AIS-related osteopenia group and the control group. Furthermore, a total RNA was extracted from isolated BMSCs to perform RNA sequencing and subsequent analysis.
RESULTS:
A lower osteogenic capacity and increased adipogenic capacity of BMSCs in AIS-related osteopenia were revealed. Differences in mRNA expression levels between the AIS-related osteopenia group and the control group were identified, including differences in the expression of LRRC17 , DCLK1 , PCDH7 , TSPAN5 , NHSL2 , and CPT1B . Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed several biological processes involved in the regulation of autophagy and mitophagy. The Western blotting results of autophagy markers in BMSCs suggested impaired autophagic activity in BMSCs in the AIS-related osteopenia group.
CONCLUSION
Our study revealed that BMSCs from AIS-related osteopenia patients have lower autophagic activity, which may be related to the lower osteogenic capacity and higher adipogenic capacity of BMSCs and consequently lead to the lower bone mass in AIS patients.
Humans
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Adolescent
;
Scoliosis/genetics*
;
Cell Differentiation/physiology*
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Osteogenesis/genetics*
;
Bone Diseases, Metabolic/genetics*
;
Kyphosis
;
Autophagy/genetics*
;
Bone Marrow Cells
;
Cells, Cultured
;
Doublecortin-Like Kinases

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