Objective To establish a human intestinal organ-on-a-chip model using a multi-array chip array to simulate the microphysiological structure of the human intestine and to investigate the impact of ionizing radiation on radiation-induced damage to human intestinal cells.Methods Caco-2 and human umbilical vein endothelial cells(HUVECs)were co-cultured in an organ chip.The cells were subjected to fluid shear stress via a precision shaker.After 7 days of dynamic culture,the morphological structure of intestinal epithelial cells and venous endothelial cells within the intestinal organ chip was examined using phase contrast microscopy,immunofluorescence staining,and confocal microscopy for three-dimensional(3D)imaging.γ-H2AX and TUNEL immunofluorescence staining were employed to assess DNA damage and apoptosis in intestinal epithelial cells two days post-irradiation.Villin immunofluorescence staining was used to evaluate villus height three days post-irradiation.EdU incorporation assay and Ki67 immunofluorescence staining were conducted to observe the effects of ionizing radiation on the proliferation of intestinal epithelial cells.Results After 7 days of dynamic culture,phase contrast microscopy and immunofluorescence staining combined with confocal 3D imaging revealed that the upper intestinal epithelial cells in the middle compartment of the chip formed a 3D intestinal villus structure,while the vascular endothelial cells in the lower compartment developed a vascular network structure.The chip was subsequently irradiated by 10 Gy X-ray.Immunofluorescence staining results indicated that the mean fluorescence intensity of γ-H2AX and TUNEL in the irradiated group was significantly higher than in the non-irradiated group 2 days after irradiation(P＜0.01),and that the proportion of EDU+and Ki67+cells in the irradiated group was significantly lower than in the non-irradiated group three days after irradiation(P＜0.05).Conclusion Caco-2 cells and HUVECs co-culture on an organ chip can generate the biomimetic structure of human intestinal villus.Ionizing radiation has been found to shorten intestinal villus,increase DNA damage and apoptosis in intestinal epithelial cells,and inhibit the proliferation of these cells.

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

Hepatocellular carcinoma （HCC） is a common tumor in the digestive tract， the formation mechanism of which remains to be fully elucidated. Although surgery， radiation， chemotherapy， targeted therapy， and immunotherapy have achieved significant results in the treatment of HCC， these methods are accompanied by a considerable number of adverse reactions and complications. In recent years， Chinese medicine has shown remarkable efficacy in the treatment of HCC， and both basic experiments and clinical studies have confirmed the effectiveness of Chinese medicine， which exerts therapeutic effects via multiple components and multiple targets. However， the pathogenesis of HCC is exceptionally complex and not fully understood， which means that studies remain to be carried out regarding the specific mechanism of Chinese medicine in preventing and treating HCC. Network pharmacology and molecular biology can be employed to decipher the mechanism of Chinese medicine in the treatment of diseases. Studies have shown that Chinese medicine can regulate various pathways such as the mitogen-activated protein kinase （MAPK）， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， Hedgehog， Wnt/β-catenin， nuclear factor-κB （NF-κB）， Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3）， and transforming growth factor-β （TGF-β）/Smad signaling pathways. Chinese medicine can exhibit its anti-HCC effects by inducing cell apoptosis， inhibiting cell proliferation and migration， and blocking the cell cycle via the above pathways. However， the specific mechanisms remain to be systematically studied. This study comprehensively reviews the regulatory effects of Chinese medicine on HCC-related signaling pathways to reveal the molecular mechanisms of Chinese medicine in the treatment of HCC. This view holds the promise of providing new targets， new perspectives， and new therapies for HCC treatment and advancing the modernization and development of Chinese medicine.

Objective To report single-center experience on CBA combined with LAAC in treat-ment of AF.Methods A retrospective study was conducted on 27 AF patients undergoing one-stop surgery of CBA combined with LAAC in Department of Cardiovascular Diseases of the People's Hospital of Liaoning Provincie from August 2020 to November 2022.The efficacy and safety of the surgery were analyzed.Results There were 22 patients(81.5％)with LAmbre and 5 patients with Watchman(18.5％,including one case of 24 mm Watchman FLX).All the patients achieved complete pulmonary vein isolation,and conversion to sinus rhythm during operation.During a mean follow-up of 30.0±9.2 months,24 patients(88.9％)maintained sinus rhythm at 12-month follow-up,and 22 patients(81.5％)maintained sinus rhythm at 24-month follow-up.TEE at 3 months after operation displayed that all the devices were in good positions and no PDL or DRT was observed.In the 11 patients undergoing cardiac enhanced CT in 12-36 months after surgery,PDL was detected in one patient(9.1％),and uncomplete endothelialisation of the device was observed in another one(9.1％)using the Watchman device.TEE at 26 months revealed one patient(3.7％)of DRT.Conclusion One-stop surgery of CBA combined with LAAC is a feasible treatment option for patients with NVAF and at high risk of stroke.

Objective:To analyze the relationship between renin-angiotensin-aldosterone system (RAAS) gene polymorphisms and susceptibility to essential hypertension (EH) in military secret service personnel.Methods:In October 2019, military secret service personnel (162 people) who were recuperating in a sanatorium from January to December 2017 were selected as the research subjects, all of whom were Han and male. The patients (79 people) who were diagnosed with EH according to the diagnostic criteria of hypertension in the "Chinese Guidelines for the Prevention and Treatment of Hypertension" (2016 Revised Edition) were the case group, and the people with normal blood pressure (83 people) were the control group. Occupational epidemiological investigation was conducted, 5 ml of fasting cubital venous blood was collected, genomic DNA was extracted by phenol-chloroform method, and RAAS gene polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism method. The distribution differences of genotype and allele frequency between groups were compared, and the relationship between genotype, allele frequency and EH was analyzed.Results:The average age of military secret service personnel was (38.2±5.3) years old, and there was no statistical significance in the average age and the age distribution over 40 years old of the case group and the control group ( P>0.05) . There were significant differences in the distribution of AGT gene M235T locus, ACE gene I/D polymorphism genotype and allele between the case group and the control group ( P<0.05) . The TT genotype with AGT gene M235T locus ( OR=3.28, 95% CI: 1.21-8.91) and DD genotype with ACE gene ( OR=2.86, 95% CI: 1.17-7.00) were risk factors for EH in military secret service personnel. Conclusion:The TT genotype of AGT gene M235T and the DD genotype of ACE gene may be the susceptibility genotypes of military secret service personnel for EH.

Objective:To analyze the relationship between renin-angiotensin-aldosterone system (RAAS) gene polymorphisms and susceptibility to essential hypertension (EH) in military secret service personnel.Methods:In October 2019, military secret service personnel (162 people) who were recuperating in a sanatorium from January to December 2017 were selected as the research subjects, all of whom were Han and male. The patients (79 people) who were diagnosed with EH according to the diagnostic criteria of hypertension in the "Chinese Guidelines for the Prevention and Treatment of Hypertension" (2016 Revised Edition) were the case group, and the people with normal blood pressure (83 people) were the control group. Occupational epidemiological investigation was conducted, 5 ml of fasting cubital venous blood was collected, genomic DNA was extracted by phenol-chloroform method, and RAAS gene polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism method. The distribution differences of genotype and allele frequency between groups were compared, and the relationship between genotype, allele frequency and EH was analyzed.Results:The average age of military secret service personnel was (38.2±5.3) years old, and there was no statistical significance in the average age and the age distribution over 40 years old of the case group and the control group ( P>0.05) . There were significant differences in the distribution of AGT gene M235T locus, ACE gene I/D polymorphism genotype and allele between the case group and the control group ( P<0.05) . The TT genotype with AGT gene M235T locus ( OR=3.28, 95% CI: 1.21-8.91) and DD genotype with ACE gene ( OR=2.86, 95% CI: 1.17-7.00) were risk factors for EH in military secret service personnel. Conclusion:The TT genotype of AGT gene M235T and the DD genotype of ACE gene may be the susceptibility genotypes of military secret service personnel for EH.

Objective:To analyze HIV testing behaviors and associated factors in men who have sex with men (MSM) and provide evidence for making intervention on promoting HIV testing.Methods:Our observational study was conducted between April and June, 2018 in Wuxi, Jiangsu province in MSM. A self-completed questionnaire was used to collect the information about their socio-demographic characteristics, sexual behaviors, acceptance of intervention services, psychological status and HIV testing behaviors. Structural equation model (SEM) was chosen to fit data and the model was modified to analyze the relationships between variables.Results:A total of 410 MSM were enrolled in our study, among whom 72.9%(299/410) were aware of the necessity of HIV testing and 69.0%(283/410) reported having HIV tests in the last year. A modified structural equation model presented good fitting results with fitting index of root mean square error of approximation (RMSEA)was 0.065, normed fit index (NFI) was 0.946, relative fit index (RFI) was 0.922, Tucker-Lewis index (TLI) was 0.968 and comparative fit index (CFI) was 0.952. Results of SEM showed that intervention service was the factor with the largest standardized total effect value of 0.57 associated with HIV testing behaviors among MSM. Among intervention services, HIV testing reminding had the biggest factor loading of 0.88. Psychological status scores played a negative role in HIV testing behaviors among MSM with the path coefficient value of -0.33.Conclusions:The awareness and testing rate of HIV among MSM population need to be further improved. While effective measures are implemented to promote HIV testing among MSM, more attention should be paid to the mental health problems of MSM.

Objective To investigate the effects and regulating mechanisms of matrix metalloproteinase 9 (MMP-9) gene silencing on aquaporin 4(AQP4) expression in astrocytes induced by oxygen-glucose deprivation.Methods Cerebral cortical astrocytes from 2 days newborn SD rats were undergone the primary culture.The ischemic cell model was established by oxygen-glucose deprivation.This experiment were divided into control group,negative control group and MMP-9 gene silencing group.The leakage rate of lactated dehydrogenase (LDH)was detected by chromatoptometry.The MMP-9 gene silencing was carried out by Lentivirus transfection.Real-time fluorescence quantitative PCR and Western blot were used to detect the expression levels of AQP4 and MMP-9.The expressions of PKA,PKC,PKG and CaMK Ⅱ were determined by Western blot.Results Compared with control group,LDH leakage rate was significantly higher in astrocytes induced by oxygen-glucose deprivation(t=13.35,P＜0.01).The expression of MMP-9 mRNA in astrocytes in MMP-9 gene silencing group(0.412±0.297) decreased significantly compared with that in negative control group(1.118 ± 0.240) (t =-4.964,P＜ 0.05).The expression of AQP4 mRNA in astrocytes in MMP-9 gene silencing group(1.002±0.082) decreased significantly compared with that in negative control group(1.442±0.066) (t=-9.886,P＜0.01).The expression of AQP4 protein in astrocytes in MMP-9 gene silencing group(0.643±0.036)decreased significantly compared with that in negative control group(1.000± 0.069)(t=-11.073,P＜0.01).The expression of PKC protein in astrocytes in MMP-9 gene silencing group (0.198±0.110)decreased significantly compared with that in negative control group (0.980± 0.232) (t =-7.218,P＜0.01).The expressions of PKA(t=0.875),PKG(t=0.818) and CaMK Ⅱ (t=0.933) protein had no statistically significant difference between negative control group and MMP-9 gene silencing group(all P＞0.05).Conclusion The permeability of astrocytes is increased by oxygen-glucose deprivation.Gene silencing MMP-9 could induce expression of AQP4 mRNA and protein decreased,and MMP-9 may regulate AQP4 expression by regulating PKC activity.

Objective To investigate the prevalence and risk factors of Toxoplasma gondii infection among key populations in Wuxi City,Jiangsu Province,so as to provide evidences for developing preventive and control interventions for T. gondii in-fection. Methods A questionnaire was designed to conduct face-to-face interviews to HIV/AIDS carriers/patients,tumor pa-tients,pregnant women and pet breeders. The blood samples were collected to test anti-Toxoplasma IgM and IgG antibodies by using ELISA method. Results Totally 404 participants were investigated,and 38 were positive in testing T. gondii antibodies (9.4%),and among these,37(97.4%)were IgG positive and 1(2.6%)was IgM positive. The multivariate logistic regression indicated that age,regularly contacting cats/dogs and being HIV positive were associated with the infection of T. gondii(OR=2.1,2.6 and 2.3,all P < 0.05). Breeding pets,being HIV positive and being tumor patients were more likely to infect T. gondii than being pregnant(OR = 1.5,8.1 and 2.8,all P < 0.05). Conclusion The high infection rate of T. gondii is observed among key populations in Wuxi City,so that the effective measurements should be taken to enhance the intervention among key popula-tions including immunocompromised people,pet breeders or people having bad dietary habits of eating raw food.