Objective To investigate the relationship between circular RNA homeodomain interacting protein ki-nase 3 (circHIPK3) and the activation of rat microglia (RM) cells.Methods In vitro, RM cells were cultured and randomized into normal and oxygen-glucose deprivation/reperfusion (OGD/R) groups, and the expression lev-el of circHIPK3 in each group was detected by RT-qPCR.The circHIPK3 lentiviral vector with puromycin resist-ance was constructed, and the overexpression (OE) group and negative control (NC) group were set up.The opti-mal multiplicity of infection (MOI) for RM cells was determined based on fluorescence expression, and puromycin was used to screen RM cells stably expressing circHIPK3 .The cells of OE and NC groups were treated with OGD/R, and the expression levels of ionized calcium binding adaptor molecule 1 (Iba-1) and eukaryotic tumor necrosis factor receptor superfamily (CD40) were detected by Western blot.The circHIPK3 translational protein potential was analyzed by the circRNAdb database, while the potential binding microRNAs on circHIPK3 were predicted by circBank and Starbase databases.Results OGD/R down-regulated circHIPK3 in RM cells (P <0.0001).The sequencing results were accurate and the lentiviral vector of circHIPK3 was constructed successfully.The optimal MOI of RM cells was 80 , puromycin at a concentration of 2μg/ml was used to screen RM cell lines stably express-ing circHIPK3 .RT-qPCR results showed that the expression level of circHIPK3 was significantly higher in the OE group compared with the NC group (P<0.01) .Western blot results revealed that the expression levels of Iba-1 and CD40 in the OE group were markedly lower than those in the NC group (P<0.05) .Protein translation analy-sis showed that circHIPK3 encoded a polypeptide of 404 amino acids with two internal ribosome entry sites (IRES) and an open reading frame (ORF) .Database analysis uncovered that circHIPK3 could target eight specific miR-Nas, namely hsa-miR-3529-5p, hsa-miR-379-5p, hsa-miR-506-3p, hsa-miR-33, hsa-miR-450b-5p, hsa-miR-551b-3p, hsa-miR-193, and hsa-miR-508-3p.Conclusion The overexpression of circHIPK3 effectively suppres-ses OGD/R-induced activation of RM cells.It has the potential to encode peptides and may act as a miRNA sponge.These findings provide a foundation for further study of circHIPK3 functions.

Objective To investigate the status of occult hepatitis B virus infection(OBI)among blood donors in Quzhou,Zhejiang,and to analyze the mutation of S region in blood donors with anti-HBc+alone and non anti-HBc+alone.Methods The OBI samples were screened by ELISA and NAT;the HBV DNA was amplified and sequenced;20 anti-HBc+alone and 25 not anti-HBc+alone samples were obtained.Results The detection rate of OBI in Quzhou was 0.10%(155/161 045),and the positive rate of anti-HBc was 74.19%(115/155).The detection rate of OBI increased with the age of blood donors(P＜0.05),but was not related to gender.The positive rate of anti-HBc+alone was 22.58%(35/155),and that of not anti-HBc alone was 51.61%(80/155).Among the 45 OBI sequencing samples,the proportion of B and C geno-type was73.33%(33/45)and 20.00%(9/45),respectively.The mutation sites of blood donors in the anti-HBc+alone group were more than those in the not anti-HBc+alone group,and the mutation rates of S114T and V168A on MHR were significantly different(P＜0.05).Conclusion The genotype of OBI infection in Quzhou is mainly type B.The mutation sites of blood donors with anti-HBc+alone are higher than those with not anti-HBc+alone,which may be more suitable as one of the OBI screening indicators.

[Objective]To study the mechanism of Quzhuo Tongbi Formula(QZTB)in improving bile acid metabolism disorder induced by high fat diet(HFD)in mice using serum metabolomics.[Methods]Male C57BL/6J mice were randomly divided into blank,HFD model and QZTB groups.Serum metabolomic analysis was performed using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-QTOF-MS)technique to screen potential biomarkers.The key targets on the enriched pathway were verified using Real-time quantitative polymerase chain reaction(RT-qPCR)method.[Results]A total of 103 potential biomarkers were screened using serum metabolomics analysis,mainly enriched in bile acid bioanabolic pathway.QZTB treatment could significantly improve the bile acid metabolism disorder induced by HFD in mice.RT-qPCR results showed that compared with blank group,the expression levels of cholesterol 7-alpha hydroxylase(CYP7A1)and cytochrome P450 family 8 subfamily B member 1(CYP8B1)in liver tissue of HFD model group were inhibited(P<0.01),and the expressions of farnitol X receptor(FXR)and small heterodimeric partner(SHP)also decreased(P<0.01,P<0.01,P<0.001,P<0.01).After QZTB treatment,the levels of key enzymes such as CYP7A1 and CYP8B1 on the classical pathway increased(P<0.01,P<0.05).FXR-SHP pathway was activated(P<0.05),thereby sterol regulatory element binding protein-1C(SREBP-1C)and fatty acid synthase(FAS)were inhibited(P<0.01,P<0.001).[Conclusion]QZTB could improve bile acid metabolism disorder induced by HFD in obese mice,which be attributed to increasing the activity of key enzymes in the classical bile acid synthase,promoting the synthesis of primary bile acid,activating FXR-SHP pathway,and further regulating bile acid biosynthesis metabolism.

Objective:To explore the incidence of hepatitis B virus (HBV) reactivation in the population with HBV associated liver cancer after receiving programmed death receptor 1 (PD-1) inhibitors combined with targeted therapy, and the prognostic differences between HBV reactivation and non reactivation populations during this treatment.Methods:A retrospective analysis was conducted on patients with primary liver cancer who received PD-1 inhibitor combined with targeted drugs treatment at the Zhongshan Hospital, Fudan University (Xiamen Branch) from January 2019 to June 2021. Clinical data such as age, sex, liver function status, cirrhosis, HBV DNA level, alpha fetoprotein, tumor stage, anti-tumor program and anti HBV program, tumor treatment response, progression free survival (PFS), and total survival (OS) were collected for t test, χ 2 test and Kaplan-Meier survival analysis. Results:A total of 66 enrolled patients were enrolled, of which 17 cases experienced HBV reactivation, with an incidence rate of 25.76%; The rates of HBV reactivation at 3 months, 6 months, 1 year, 2 years, and 3 years were 6.06%(4/66), 12.12%(8/66), 19.70%(13/66), 22.73%(15/66), and 25.76%(17/66), respectively. There was no significant difference between the HBV reactivation group and the non HBV reactivation group in age, sex, liver function status, cirrhosis, HBV DNA level, alpha fetoprotein, tumor stage, anti-tumor and anti HBV programs, objective response rate (ORR) and disease control rate (DCR) (all P>0.05). However, the PFS and OS of the HBV reactivation group were significantly lower than those of the non HBV reactivation group, at 4.00 months vs 8.50 months ( P=0.002) and 12.90 months vs 19.77 months ( P=0.014), respectively. Conclusions:Patients with primary live cancer who receive PD-1 inhibitor combined with targeted therapy are at risk of HBV reactivation, and those who experience HBV reactivation have significantly poorer tumor progression and survival prognosis compared with non HBV reactivated patients.

Objective To explore the impact of SARS-CoV-2-positive donors on the prognosis of heart transplant recipients. Methods The Medline, EMbase, CENTRAL, CNKI, Wanfang Data, VIP and China Biology Medicine from inception to May 2023 were searched by computer for studies about impact of SARS-CoV-2-positive donors on the prognosis of heart transplant recipients. The data were extracted from all the relevant literatures, and the quality of the data was assessed using the Newcastle-Ottawa Scale (NOS). All statistical analyses were conducted by the Stata 11.0 software. Results A total of 10 studies (NOS score ranging from 5 to 9 points) involving 643 patients were enrolled. The pooled results demonstrated that the pooled mortality of heart transplant recipients from SARS-CoV-2-positive donors was 4% (95%CI 2% to 5%). And the incidence of composite outcome, regarding graft failure, rejection and death as poor prognosis, was 7% (95%CI 5% to 9%). Besides, compared with recipients from SARS-CoV-2-negative donors, the pooled odds ratio (OR) value of death of SARS-CoV-2-positive donors was 0.68 (95%CI 0.38 to 1.22, Z=1.28, P=0.200). The pooled OR value of rejection rate was 0.41 (95%CI 0.27 to 0.64, Z=3.97, P<0.005). For the composite outcome, the pooled OR value was 0.50 (95%CI 0.37 to 0.69, Z=4.30, P<0.005). In addition, there was no statistical difference in the length of hospital stay between heart transplant recipients from SARS-CoV-2-positive donors and negative donors (SMD=–0.03, 95%CI –0.22 to 0.15, Z=0.36, P=0.720). Conclusion The application of heart from SARS-CoV-2-positive donor for transplantation is safe and feasible. However, further prospective studies with longer follow-up are still needed to verify its impact on long-term outcomes.

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.

Objective To investigate expression levels and clinical significance of annexin A2 and S100 calcium binding protein A10 (S100A10) in cerebrospinal fluid of patients with secondary intracranial infection after craniocerebral surgery. Methods A total of 120 patients with secondary intracranial infection after craniocerebral surgery were selected as test group, while 120 patients with no infection after craniocerebral surgery in the same period were selected as control group. The levels of Annexin A2 and S100A10 in cerebrospinal fluid were detected by enzyme-linked immunosorbent assay(ELISA). Pearson correlation analysis was applied to analyze the correlations of Annexin A2 and S100A10 with clinical indicators. Logistic regression analysis was applied to analyze the influencing factors of secondary intracranial infection after craniocerebral surgery. Receiver operator characteristic (ROC) curve was applied to analyze the predictive value of Annexin A2 and S100A10 levels for the occurrence of secondary intracranial infection after craniocerebral surgery. Results The proportions of diabetes and cerebrospinal fluid leakages, blood l actate dehydrogenase (LDH), cerebrospinal fluid Annexin A2 and S100A10 levels in the test group were higher than those in the control group (P < 0.05). Annexin A2 in cerebrospinal fluid of patients with intracranial infection was positively correlated with S100A10, LDH, and S100A10 level was positively correlated with LDH (P < 0.05). Multivariate Logistic regression analysis showed that diabetes, cerebrospinal fluid leakage, blood LDH, Annexin A2 and S100A10 in cerebrospinal fluid were independent influencing factors for secondary intracranial infection after craniocerebral surgery (P < 0.05). ROC results showed that the AUCs of Annexin A2 level or S100A10 level alone in cerebrospinal fluid and their combined prediction for secondary intracranial infection after craniocerebral surgery was 0.788, 0.768 and 0.865 respectively, the AUC of combined prediction was larger than that of the single prediction (P < 0.05). Conclusion The expression levels of Annexin A2 and S100A10 are increased in cerebrospinal fluid of patients with secondary intracranial infection after craniocerebral surgery, which are independent influencing factors for secondary intracranial infection after craniocerebral surgery. Their combination has higher predictive value for secondary intracranial infection after craniocerebral surgery.

Objective:To investigate the correlation between triglyceride-glucose (TyG) index and high on-treatment platelet reactivity (HTPR) during clopidogrel treatment in patients with ischemic stroke.Methods:Patients with ischemic stroke who received maintenance dose of clopidogrel (75 mg/d) in the Department of Neurology, Guangdong Provincial Hospital of Chinese Medicine from January 2017 to March 2021 were retrospectively included. The highest quartile (Q4) of the TyG index was defined as insulin resistance. Platelet reactivity was assessed by thromboelastogram and clopidogrel HTPR was defined as the clot strength induced by adenosine diphosphate (MA ADP) >47 mm. Multivariate regression model was used to analyze the independent correlation between TyG index and platelet reactivity. Results:A total of 83 patients were included. The TyG index showed a linear correlation with MA ADP. The patients were divided into 4 groups according to the quartile of TyG index. The incidence of clopidogrel HTPR increased significantly with the increase of the quartile of the TyG index ( Ptrend=0.017). Multivariate analysis showed that there was a significant independent correlation between insulin resistance and clopidogrel HTPR (odds ratio 4.597, 95% confidence interval 1.285-16.446; P=0.019). Conclusions:In patients with ischemic stroke treated with clopidogrel, the incidence of clopidogrel HTPR gradually increases with the increase of the quartile of the TyG index. The insulin resistance assessed by the TyG index is independently associated with clopidogrel HTPR.

Humans ; Hypertension, Pulmonary/surgery* ; Learning Curve ; Angioplasty, Balloon ; Pulmonary Embolism/therapy* ; Chronic Disease ; Pulmonary Artery/surgery*