Purpose: Our previous study showed that circulating tumor cell (CTC) count combined with gene mutation detection might help differentiate benign and malignant pulmonary nodules (PNs). Herein, we aimed to expand the study cohort and conduct further sequencing analysis. Materials and Methods: Patients with PNs were included, and CTCs were identified before operation. Low-coverage whole-genome sequencing (LC-WGS) and lung cancer-related targeted gene sequencing were performed on CTCs. The diagnostic efficacy was evaluated by receiver operating characteristic (ROC) curve. The differences in CTC counts among subgroups classified by demographic–clinical characteristics were analyzed. LC-WGS–based copy number variation (CNV) analysis and targeted gene mutation analysis were conducted. Results: A total of 172 patients were included. CTC count of 2.5 was identified by the ROC curves as the optimal diagnostic cutoff. The sensitivity and specificity of CTC count for differentiating benign and malignant PNs were 54.2% and 78.6%, respectively. The diagnostic sensitivity and specificity of combined CTC count, radiological nodule type, and any malignant imaging features were 84.7% and 71.4%, respectively. The CTC counts were significantly greater in patients with aggressive tumors, later stage, and spread through air spaces. CTCs from malignant cases had more CNVs than those from benign cases. Conclusion CTC count can be used in identifying malignant PNs. The diagnostic efficacy can be improved if combined with computed tomography imaging characteristics. Further CNV analysis might help differential diagnosis. Greater CTC count might suggest more aggressive tumors. CTC detection can provide important information and guidance for subsequent management of PNs.

Objective To systematically review the efficacy and safety of the tislelizumab combined with anlotinib regimen for advanced non-small-cell lung cancer(NSCLC)patients.Methods PubMed,Embase,Cochrane Library,Web of Science,CNKI,WanFang Data databases were electronically searched to collect clinical studies on the combination of trastuzumab and anlotinib in the treatment of advanced NSCLC patients from inception to August 10,2024.Two reviewers independently screened literature,extracted data and assessed the risk of bias of the included studies.Meta-analysis was then performed by using RevMan 5.4.1 software.Results A total of 6 articles of 468 patients were included,involving 4 randomized controlled trials and 2 prospective cohort studies.Meta-analysis showed that the tislelizumab combined with anlotinib group had higher ORR[OR=2.53,95％CI(1.62,3.93),P＜0.001]and DCR[OR=4.45.95％CI(2.43,8.15),P＜0.001]than the anlotinib group.The CYFRA21-1 level in the combination group was significantly lower than that in the anlotinib group[SMD=-1.07,95％CI(-1.63,-0.51),P＜0.001].For safety,there were no significant differences in the incidence of adverse reaction of leukopenia[OR=0.91,95％CI(0.39,2.14),P=0.83],liver/kidney dysfunction[OR=1.16,95％CI(0.39,3.44)P=0.78].The descriptive analysis results indicated that there was no statistically significant difference in CEA levels between the two groups before treatment in each study(P＞0.05),and the CEA levels in the combination therapy group were lower than those in the anlotinib group after treatment(P＜0.05).Conclusion Compared to anlotinib alone,tislelizumab combined with anlotinib improves ORR,DCR,and reduces tumor markers in NSCLC patients,with comparable incidence of adverse reactions.Due to the limited quality and quantity of the included studies,more high quality studies are needed to verify the above conclusion.

Objective To systematically review the efficacy and safety of the tislelizumab combined with anlotinib regimen for advanced non-small-cell lung cancer(NSCLC)patients.Methods PubMed,Embase,Cochrane Library,Web of Science,CNKI,WanFang Data databases were electronically searched to collect clinical studies on the combination of trastuzumab and anlotinib in the treatment of advanced NSCLC patients from inception to August 10,2024.Two reviewers independently screened literature,extracted data and assessed the risk of bias of the included studies.Meta-analysis was then performed by using RevMan 5.4.1 software.Results A total of 6 articles of 468 patients were included,involving 4 randomized controlled trials and 2 prospective cohort studies.Meta-analysis showed that the tislelizumab combined with anlotinib group had higher ORR[OR=2.53,95％CI(1.62,3.93),P＜0.001]and DCR[OR=4.45.95％CI(2.43,8.15),P＜0.001]than the anlotinib group.The CYFRA21-1 level in the combination group was significantly lower than that in the anlotinib group[SMD=-1.07,95％CI(-1.63,-0.51),P＜0.001].For safety,there were no significant differences in the incidence of adverse reaction of leukopenia[OR=0.91,95％CI(0.39,2.14),P=0.83],liver/kidney dysfunction[OR=1.16,95％CI(0.39,3.44)P=0.78].The descriptive analysis results indicated that there was no statistically significant difference in CEA levels between the two groups before treatment in each study(P＞0.05),and the CEA levels in the combination therapy group were lower than those in the anlotinib group after treatment(P＜0.05).Conclusion Compared to anlotinib alone,tislelizumab combined with anlotinib improves ORR,DCR,and reduces tumor markers in NSCLC patients,with comparable incidence of adverse reactions.Due to the limited quality and quantity of the included studies,more high quality studies are needed to verify the above conclusion.

Pulmonary hypertension (PH) is an insidious pulmonary vasculopathy with high mortality and morbidity and its underlying pathogenesis is still poorly delineated. The hyperproliferation and apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs) contributes to pulmonary vascular remodeling in pulmonary hypertension, which is closely linked to the downregulation of fork-head box transcriptional factor O1 (FoxO1) and apoptotic protein caspase 3 (Cas-3). Here, PA-targeted co-delivery of a FoxO1 stimulus (paclitaxel, PTX) and Cas-3 was exploited to alleviate monocrotaline-induced pulmonary hypertension. The co-delivery system is prepared by loading the active protein on paclitaxel-crystal nanoparticles, followed by a glucuronic acid coating to target the glucose transporter-1 on the PASMCs. The co-loaded system (170 nm) circulates in the blood over time, accumulates in the lung, effectively targets the PAs, and profoundly regresses the remodeling of pulmonary arteries and improves hemodynamics, leading to a decrease in pulmonary arterial pressure and Fulton's index. Our mechanistic studies suggest that the targeted co-delivery system alleviates experimental pulmonary hypertension primarily via the regression of PASMC proliferation by inhibiting cell cycle progression and promoting apoptosis. Taken together, this targeted co-delivery approach offers a promising avenue to target PAs and cure the intractable vasculopathy in pulmonary hypertension.

Objective:To evaluate the influence of cytomegalovirus (CMV) infection on T cell senescence and cardiovascular disease (CVD) in maintenance hemodialysis (MHD) patients.Methods:It was a single center cross sectional study. Patients aged over 18 years old and received hemodialysis for at least 6 months at the Blood Purification Centre of the Department of Nephrology of Zhongshan Hospital Affiliated to Fudan University from January 2021 to April 2021 were enrolled. Demographic, hematological, nutritional and inflammatory markers were obtained. Anti-CMV-IgM and IgG antibodies were detected using the Roche Elecsys assay. CD28 - T cell was evaluated by flow cytometry. Mann-Whitney U test or Kruskal-Wallis H test was used for anti-CMV-IgG comparison among groups. Spearman correlation and linear regression were used to assess the relationship between anti-CMV-IgG and CD28 - T cell compartment. Logistic regression was used to assess the relationship between anti-CMV-IgG and CVD. Results:A total of 438 MHD patients (270 men and 168 women) were enrolled in the study. The median age was 62 (51, 70) years. The median time on hemodialysis was 57 (21, 100) months. The primary diseases included chronic glomerulonephritis [213 cases (48.6%)], diabetic nephropathy [82 cases (18.7%)], polycystic kidney disease [34 cases (7.8%)], hypertensive renal disease [34 cases (7.8%)], etc. Of these patients, 430 (98.2%) were seropositive for anti-CMV-IgG, 206 (47.0%) had anti-CMV-IgG titers exceeding the upper limit of 500 U/ml. Patients aged over 70 years old were 100% seropositive for anti-CMV-IgG. Patients on HD for more than 5 years had a higher seropositive rate of 99.1% than those with shorter HD duration, although these results were not statistically significant. Spearman correlation analysis showed that the anti-CMV-IgG titers in MHD patients were positively correlated with the proportion of CD4 + CD28 - T cells and CD8 + CD28 - T cells ( r=0.316, P<0.001; r=0.272, P<0.001). Multiple linear regression analysis showed that after adjusting for age and gender, lg[CD4 + CD28 - T cells(%)] and lg[CD8 + CD28 - T cells(%)] were positively correlated with lg[anti-CMV-IgG titers (U/ml)], respectively ( β=0.455, t=8.315, P<0.001; β=0.412, t=7.282, P<0.001). In analyzing the relationship between anti-CMV-IgG titers and CVD, patients were divided into six groups according to age and anti-CMV-IgG level. Group 1 included young patients with a lower anti-CMV-IgG titers (age ≤55 years old, anti-CMV-IgG <400 U/ml); Group 2 included young patients with a higher anti-CMV-IgG titers (age≤55 years old, anti-CMV-IgG ≥400 U/ml); Group 3 included middle-aged patients with a lower anti-CMV-IgG titers (5565 years old, anti-CMV-IgG<400 U/ml); Group 6 included old aged patients with a higher anti-CMV-IgG titers (age>65 years old, anti-CMV-IgG≥400 U/ml). The incidence of CVD was significantly different among the six groups ( χ2=18.780, P=0.002) and patients aged over 55 years old with higher anti-CMV-IgG level had a higher CVD incidence compared with group 1 ( P<0.05). Multivariate logistic regression analysis showed that increased age ( OR=1.020, 95% CI 1.002-1.038, P=0.033), male ( OR=1.855, 95% CI 1.161-2.965, P=0.010), history of diabetes ( OR=1.867, 95% CI 1.145-3.046, P=0.012), increased lg[NT-proBNP(μg/L)] ( OR=2.848, 95% CI 1.816-4.467, P<0.001) and lg[anti-CMV-IgG (U/ml)] ( OR=3.183, 95% CI 1.582- 6.405, P=0.001) were independent factors associated with CVD in MHD patients. Conclusions:CMV infection is extremely common in MHD patients. Increased anti-CMV-IgG is related to T cell senescence and CVD complications in MHD patients.

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

Objective:To evaluate the relationship between chemokine CXC-ligand 16 (CXCL16) and natural killer T cells during renal fibrosis in mice with acute kidney injury (AKI).Methods:Twenty-four healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), AKI group, control+ rCXCL16 group (group C-rCXCL16) and AKI+ rCXCL16 group.In AKI-rCXCL16 and AKI groups, folic acid 250 mg/kg was intraperitoneally injected to induce AKI in anesthetized mice, and rCXCL16 0.1 mg/kg and the equal volume of solution were intraperitoneally injected, respectively, at 3, 6, 9 and 12 days after injection of folic acid.The equal volume of solution and rCXCL16 were intraperitoneally injected at the corresponding time points in group C and group C-rCXCL16, respectively.The orbital blood samples were taken on day 14 after injection of folic acid for determination of the serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The renal tissues were obtained for measurement of the renal fibrosis size (using Sirius red staining and Masson staining), for determination of the expression of fibronectin (FN), collagen-Ⅲ (Col-Ⅲ) and α-smooth muscle actin (α-SMA) (by immunofluorescence) and expression of interleukin-4 (IL-4), mannose receptor (CD206) and arginase 1 (Arg-1) mRNA (by real-time polymerase chain reaction), and for evaluation of the ratio of CD1d Tetramer + -IL-4 + cells (by flow cytometry). Results:Compared with group C, the serum BUN and Cr concentrations were significantly increased, the renal fibrosis size was increased, the expression of IL-4, CD206, Arg-1 mRNA, FN, Col-Ⅲ and α-SMA was up-regulated, and the ratio of CD1d Tetramer + -IL-4 + cells was increased in AKI and AKI-rCXCL16 groups ( P<0.05), and no significant change was found in the parameters mentioned above in group C-rCXCL16 ( P>0.05). Compared with group AKI, the serum BUN and Cr concentrations were significantly increased, the renal fibrosis size was increased, the expression of IL-4, CD206, Arg-1 mRNA, FN, Col-Ⅲ and α-SMA was up-regulated, and the ratio of CD1d Tetramer + -IL-4 + cells was increased in group AKI-rCXCL16 ( P<0.05). Conclusion:The mechanism by which CXCL16 is involved in the process of renal fibrosis is related to the recruitment of natural killer T cells secreting IL-4 which regulates macrophage M2 polarization in mice with AKI.

Objective To explore the dosimetric characteristics of single photon emission computed tomography(SPECT)combined with bone marrow sparing intensity modulated radiotherapy in patients with cervical cancer.Methods Twenty patients with cervical cancer after hysterectomy were performed on 99Tc thiocolloid SPECT bone marrow imaging to determine the active bone marrow in the pelvis.SPECT image combined with the simulation CT was used as the primary planning data set.Two plans for bone marrow sparing modulated radiotherapy were generated according to the bone marrow volume defined by SPECT and outline of the pelvis(SPECT-IMRT planning and BMS-IMRT planning).Furthermore,the target section and the dosimetric distribution in organs at risk(bone marrow,small intestine,rectum and bladder)in the two plannings were compared.The prescribed dose to the PTV was 45 Gy.Results The volume of the bone marrow in SPECT-IMRT planning and BMS-IMRT planning was(238.15 ±36.82)and(1 100.61 ±109.92)cm3,respectively(t＝33.273,P＜0.05).The average bone marrow volume of SPECT-IMRT planning in high-dose radiation region(V30,V40and V45)was decreased by 6.9％,5.7％and 2.6％respectively compared with BMS-IMRT planning(t ＝3.540,3.4261,3.448,P＜0.05),but there was no statistical significant difference between the two plannings in the dose volume of low-dose radiation(V10and V20,P＞0.05).The PTV coverage rate and exposure dose of other organs at risk(small intestine,rectum and bladder)in the two plannings were not significantly different(P＞0.05).Conclusions SPECT bone marrow imaging could clearly show the range of active bone marrow on CT scan images.Compared with BMS-IMRT,SPECT-IMRT could further reduce the dose volume of high-dose radiation(V30,V40and V45).Yet the clinical significance of this advantage also needs to be confirmed by large-scale clinical studies.

Objective To observe the changes of subfoveal choroidal thickness (SFCT) in diabetic patients with non-proliferative diabetic retinopathy (NPDR) and clinically significant macular edema(CSME),and then investigate diabetic SFCT and the relationship of diabetic CT with diabetic retinopathy (DR).Methods The patients were divided into 2 groups according to clinical guidelines of DR in China in 2014,including NPDR CSME + group(21 eyes) and NPDR CSME-group(36 eyes).All patients were underwent best corrected visual acuity (BCVA),intraocular pressure,axial length,slit lamp microscope,indirect ophthalmoscope,EDI-OCT examination,and mean arterial blood pressure measurement.The differences of BCVA,central retinal thickness (CRT) and SFCT between NPDR CSME + group and NPDR CSME-group were studied as well by means of using SPSS 18.0 for data statistics.Results There was no significant difference in gender,age,eye axis and intraocular pressure between NPDR CSME + group and NPDR CSME-group (all P ＞ 0.05).There was significant difference in BCVA between the two groups (P =0.001).The mean SFCT were (328.24 ± 101.92) μm in the NPDR CSME + group and (235.31 ± 66.98) μm in the NPDR CSME-group,and the difference was statistically significant (t =4.156,P=0.000).And plotting changes in CRT against changes in SFCT in patients with CSME revealed a positive correlation(r =0.473,P =0.000).Conclusion SFCT in NPDR CSME + patient is thicker than that in NPDR CSME-patient.And plotting changes in CRT against changes in SFCT in patients with CSME reveals a positive correlation.

Objective To clone and identify the heat shock factors(HSFs)of Schistosoma japonicum and analyze its molec?ular structure and alternative splicing pattern. Methods The New Zealand rabbits were infected with the cercariae of Schistoso?ma japonicum and were killed and dissected 42 days post?infection,and the adult worms of S. japonicum and the livers of the rabbits were harvested. Then,the total RNA was extracted by using Trizol reagent. The Sj?hsf open reading frame(ORF)and the alternative splicing fragments were amplified by RT?PCR from the female,male and egg samples,then cloned and verified by enzyme digestion and sequencing. DNAMAN 8.0,InterPro,Mega 6 combined with the Internet databases were utilized to clarify the gene structure,functional domains,alternative splicing pattern,and the homology and phylogenetic tree of HSFs. Re?sults Sj?hsf ORF and the alternative splicing fragments were amplified from the female,male and egg samples of S. japonicum by RT?PCR. After cloning,the positive recombinant plasmids pBSjHSFf?F,pBSjHSFf?M,pBSjHSFf?E containing Sj?hsf ORF, pBSjHSFs?F,pBSjHSFs?M,pBSjHSFs?E with Sj?hsf alternative splicing fragments were identified by enzyme digestion and se?quencing. Three alternative splicing Sj?hsf isoforms were observed through sequence analysis:Sj?hsf?isoform1(2 050 bp),Sj?hsf ?isoform2(2 086 bp)and Sj?hsf?isoform3(2 111 bp);the GenBank accession numbers were KU954546,KX119143 and KX119144,respectively. All the three isoforms located in the same Contig SJC_S000780 of S. japonicum genome and all ex?pressed at female,male and egg stages,but Sj?hsf?isoform1 with a high?level expression. Sj?HSF?isoform1(671 aa)and Sj?HSF?isoform2(683 aa)had DBD(DNA binding domain),HR?A/B and HR?C domains,while Sj?HSF?isoform3(282 aa)stopped in advance without HR?C domain. Phylogenetic tree analysis of HSFs illustrated that Sj?HSFs belonged to HSF1 family,with a close phylogenetic relationship to Sm?HSFs. Conclusions There are three alternative splicing isoforms of Sj?HSF existing in the female,male and egg stages of S. japonicum,but Sj?HSF?isoform1 expresses in a high?level. This study lays the foundation for further study on molecular mechanisms of Sj?HSFs in regulating the heat shock response system.