Background and purpose:Immune checkpoint blockade(ICB)has become a promising strategy for treating cervical cancer(CC),but terminal T cell depletion still limits the therapeutic efficacy of ICB.The deletion of sorting nexin-9(SNX9)can inhibit thymocyte selection-associated high mobility group box protein(TOX),alleviate T cell exhaustion,and provide new ideas for preventing T cell exhaustion and enhancing the efficacy of cancer immunotherapy.Therefore,this study aimed to explore the immune escape mechanism mediated by the depletion of CD8+T cells induced by the SNX9/TOX signaling pathway in the CC microenvironment.Methods:Fifty-four peripheral blood samples were collected,including 18 from CC patients,18 from patients with high-grade squamous intraepithelial lesions(HSIL)and 18 from subjects with normal cervix.In addition,the study collected 153 pairs of CC and adjacent tissues from patients who received operation in our hospital for the first time.Clinicopathological features,tumor stages,follow-up records and other relevant clinicopathological data of CC patients were obtained from hospital records.The research was approved by the ethics committee of Changsha Fourth Hospital(approval number:20220206).A total of 24 mice were randomly assigned to the following four groups:immunoglobulin G(IgG)group,Anti-SNX9 group,Anti-programmed death-1(PD-1)group and Anti-SNX9+Anti-PD-1 group,with 6 mice in each group.Each group received intraperitoneal injection of blocking antibody and isotype control treatment respectively.ELISpot was used to detect the ability of CD8+T cells to secrete tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ).The expressions of TOX and SNX9 in cervical cancer tissues were detected by western blot and immunohistochemistry.Results:The expressions of SNX9 and TOX mRNA in peripheral blood mononuclear cell(PBMC)of CC patients were higher compared with HSIL and normal controls(P＜0.05).The positive cell level of SNX9 and TOX immunohistochemical score were higher in CC tissue than in adjacent tissues(t=18.63,21.10,P＜0.001).The high expression of SNX9 in CC was related to low differentiation/undifferentiation,tumor size,parauterine infiltration,vaginal infiltration,late FIGO stage and pelvic lymph node metastasis(P＜0.05).Compared with the low expression group of SNX9,the overall survival time of CC patients in the high expression group of SNX9 was shorter(P＜0.05).The percentage of CD8+SNX9+T cells was significantly higher in CC patients than in normal controls and HSIL patients(P＜0.05).The ability of CD8+SNX9+T cells to secrete cytokines(TNF-α and IFN-γ)was significantly lower compared with CD8+SNX9-T cells(P＜0.05).Compared with the Anti-SNX9 group,the growth and proliferation of cervical tumor,the expression of SNX9 and TOX protein in tumor tissue in the Anti-SNX9+Anti-PD-1 group further decreased(P＜0.05),and the level of infiltrating CD8+T lymphocytes in tumor tissue and the ability of CD8+T lymphocytes to secrete functional factors TNF-α and IFN-γ further increased(P＜0.05).Conclusion:SNX9/TOX signaling pathway exhibits enhanced activity in patients with cervical cancer and mouse models,and is related to immunosuppression.Targeting SNX9/TOX signaling pathway may be a potential therapeutic strategy for CC.

Background and purpose:Immune checkpoint blockade(ICB)has become a promising strategy for treating cervical cancer(CC),but terminal T cell depletion still limits the therapeutic efficacy of ICB.The deletion of sorting nexin-9(SNX9)can inhibit thymocyte selection-associated high mobility group box protein(TOX),alleviate T cell exhaustion,and provide new ideas for preventing T cell exhaustion and enhancing the efficacy of cancer immunotherapy.Therefore,this study aimed to explore the immune escape mechanism mediated by the depletion of CD8+T cells induced by the SNX9/TOX signaling pathway in the CC microenvironment.Methods:Fifty-four peripheral blood samples were collected,including 18 from CC patients,18 from patients with high-grade squamous intraepithelial lesions(HSIL)and 18 from subjects with normal cervix.In addition,the study collected 153 pairs of CC and adjacent tissues from patients who received operation in our hospital for the first time.Clinicopathological features,tumor stages,follow-up records and other relevant clinicopathological data of CC patients were obtained from hospital records.The research was approved by the ethics committee of Changsha Fourth Hospital(approval number:20220206).A total of 24 mice were randomly assigned to the following four groups:immunoglobulin G(IgG)group,Anti-SNX9 group,Anti-programmed death-1(PD-1)group and Anti-SNX9+Anti-PD-1 group,with 6 mice in each group.Each group received intraperitoneal injection of blocking antibody and isotype control treatment respectively.ELISpot was used to detect the ability of CD8+T cells to secrete tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ).The expressions of TOX and SNX9 in cervical cancer tissues were detected by western blot and immunohistochemistry.Results:The expressions of SNX9 and TOX mRNA in peripheral blood mononuclear cell(PBMC)of CC patients were higher compared with HSIL and normal controls(P＜0.05).The positive cell level of SNX9 and TOX immunohistochemical score were higher in CC tissue than in adjacent tissues(t=18.63,21.10,P＜0.001).The high expression of SNX9 in CC was related to low differentiation/undifferentiation,tumor size,parauterine infiltration,vaginal infiltration,late FIGO stage and pelvic lymph node metastasis(P＜0.05).Compared with the low expression group of SNX9,the overall survival time of CC patients in the high expression group of SNX9 was shorter(P＜0.05).The percentage of CD8+SNX9+T cells was significantly higher in CC patients than in normal controls and HSIL patients(P＜0.05).The ability of CD8+SNX9+T cells to secrete cytokines(TNF-α and IFN-γ)was significantly lower compared with CD8+SNX9-T cells(P＜0.05).Compared with the Anti-SNX9 group,the growth and proliferation of cervical tumor,the expression of SNX9 and TOX protein in tumor tissue in the Anti-SNX9+Anti-PD-1 group further decreased(P＜0.05),and the level of infiltrating CD8+T lymphocytes in tumor tissue and the ability of CD8+T lymphocytes to secrete functional factors TNF-α and IFN-γ further increased(P＜0.05).Conclusion:SNX9/TOX signaling pathway exhibits enhanced activity in patients with cervical cancer and mouse models,and is related to immunosuppression.Targeting SNX9/TOX signaling pathway may be a potential therapeutic strategy for CC.

Objectives:To investigate the clinical value of contrast echocardiography in the detection of right heart space-occupying lesions. Methods:The imaging and clinical data of 39 patients with right heart space-occupying lesions confirmed by surgery,pathology and clinical treatment in Yanbian University Hospital from January 2019 to January 2023 were retrospectively analyzed.All patients underwent contrast echocardiography and cardiac magnetic resonance(CMR)examinations.The contrast echocardiography features of right heart neoplastic from non-neoplastic occupying lesions were summarized.The diagnostic efficacy of contrast echocardiography and CMR in right heart space-occupying lesions was compared. Results:Among the 39 patients,12 patients(30.8%)were confirmed as thrombus and vegetation,6 patients(15.4%)as benign tumors and 21 patients(53.8%)as malignant tumors.There were 8,4,4 and 17 patients of thrombus,vegetation,benign tumor and malignant tumors located in right atrium,respectively.Contrast echocardiography was used to diagnose thrombus and vegetation in 12 patients(30.8%),benign tumors in 5 patients(12.8%),malignant tumors in 22 patients(56.4%).CMR was used to diagnose thrombus and vegetation in 18 patients(46.2%),benign tumors in 5 patients(12.8%),malignant tumors in 16 patients(41.0%).The Kappa coefficient between contrast echocardiography and clinical diagnosis was 0.956(P<0.001).The Kappa coefficient between CMR and clinical diagnosis was 0.754(P<0.001). Conclusions:Contrast echocardiography can dynamically observe the degree of vascularization in the lesion tissue in real-time,accurately distinguishing the nature of the right heart space-occupying lesions.

Chronic alcohol consumption causes liver steatosis, cell death, and inflammation. Melatonin (MLT) is reported to alleviate alcoholic liver disease (ALD)-induced injury. However, its direct regulating targets in hepatocytes are not fully understood. In the current study, a cell-based screening model and a chronic ethanol-fed mice ALD model were used to test the protective mechanisms of MLT. MLT ameliorated ethanol-induced hepatocyte injury in both cell and animal models (optimal doses of 10 μmol/L and 5 mg/kg, respectively), including lowered liver steatosis, cell death, and inflammation. RNA-seq analysis and loss-of-function studies in AML-12 cells revealed that telomerase reverse transcriptase (TERT) was a key downstream effector of MLT. Biophysical assay found that epidermal growth factor receptor (EGFR) on the hepatocyte surface was a direct binding and regulating target of MLT. Liver specific knock-down of Tert or Egfr in the ALD mice model impaired MLT-mediated liver protection, partly through the regulation of nuclear brahma-related gene-1 (BRG1). Long-term administration (90 days) of MLT in healthy mice did not cause evident adverse effect. In conclusion, MLT is an efficacious and safe agent for ALD alleviation. Its direct regulating target in hepatocytes is EGFR and downstream BRG1-TERT axis. MLT might be used as a complimentary agent for alcoholics.

Objective:To investigate the sensitivity of newborn screening for neonatal intrahepatic cholestasis caused by Citrin deficiency (NICCD) based on tandem mass spectrometry and the carrying rate of known pathogenic variants of SLC25A13 in Guangzhou population. Methods:A total of 124 250 neonates born in Guangzhou from January 1, 2015 to December 31, 2018 were performed newborn screening for NICCD by tandem mass spectrometry technology. SLC25 A13 gene mutation analysis was performed to diagnose patients with suspected NICCD.The carrying rate of known pathogenic variants of the SLC25 A13 gene in the whole exon sequencing results of 2 395 healthy children in Guangzhou was retrospective analyzed. Results:Among the 124 250 screened neonates, 31 cases were screened positive for NICCD and one of them was confirmed.Three false negative patients with NICCD were found in this cohort.NICCD screening sensitivity was 25%(1/4 cases). All of the four patients were homozygous for c. 851_854del of SLC25A13.Among 2 395 controls, 60 cases were detected heterozygous variant of SLC25A13, including 8 kinds of reported pathogenic variants.The carrying rate of pathogenic alleles was 1/40 (60/2 395 cases). The estimated prevalence of citrin deficiency was about 1/6 400.The most common variant was c. 851_854del (56.7%, 34/60 cases), and the second was c. 790G>A (23.3%, 14/60 cases). The controversial variant c. 2T>C was detected in 113 children with heterozygous and 2 cases with homozygous and the carrying rate of c. 2T>C was 1/20(117/2 395 cases). Conclusions:The carrying rate of pathogenic variants of SLC25A13 and the estimated prevalence of Citrin deficiency in Guangzhou population are high.The sensitivity of newborn screening for NICCD by tandem mass spectrometry is limited.Even if the negative results for screening of multiple genetic and metabolic diseases by tandem mass spectrometry, it is recommended to recheck blood for newborns or infants with delayed jaundice to avoid missed diagnosis.

Objective To investigate the epidemiology characteristics of crawfish related rhabdomyolysis (RM) in Nanjing, 2016.Methods Outpatient and inpatient electronic medical system of 21 hospitals in Nanjing during 2016 were retrospectively searched, and all the patients diagnosed with RM were selected. The patients with none crayfish-related RM was excluded. The epidemiology characteristics were depicted. The geographic information system (GIS) was used to collect, manage and analyze the spatial data, to visualize it, to analyze the spatial distribution features of the disease, and to explore the cause of disease prediction. GeoDa 1.8 software was used to analyze the global and local spatial auto-correlation.Results A total of 1183 patients with crawfish related RM were initially screened, excluding 59 patients with RM caused by trauma, severe exercise, heat stroke, myositis, poisoning, drugs, and genetic diseases, and 1124 patients were enrolled. The proportion of men was 36.48% (410/1124) with an incidence of 12.54/100 thousands; while of women was 63.52% (714/1124) with an incidence of 21.86/100 thousands. The median age at onset was 34 (28, 43) years. From July to August, the incidence of crawfish related RM was the highest, accounting for 96.53% of the total number of cases. The top four incidence areas were Pukou (41.54/100 thousands), Jianye (25.94/100 thousands), Qixia (25.73/100 thousands), Gulou (25.04/100 thousands), all of which were adjacent to the Yangtze River. Global spatial autocorrelation analysis showed: MoranI = 0.427,Z = 2.646,P = 0.003, suggesting that the crawfish related RM had positive spatial autocorrelation. The results showed that the spatial structure of crawfish related RM existed in Nanjing in 2016. Local spatial autocorrelation analysis showed that the high-high concentration areas were Pukou, Jianye and Liuhe. The incidences of above three areas which were the Nanjing section of the lower reaches of the Yangtze River flowed through the region and surrounding areas were higher than the overall incidence of Nanjing.Conclusion The prevalence of crawfish related RM in Nanjing during 2016 had an obvious region-concentrated character and global spatial autocorrelation with the high prevalent regions mainly concentrated in the urban areas adjacent to the Yangtze River.

[Objective]To compare the differences of neuromyelitis optica spectrum disorder(NMOSD)and multiple sclerosis (MS) on pregnancy ,and analyze the mutual impact of pregnancy on the diseases.[Methods]Prospectively collected clinical information of 235 NMOSD patients and 125 MS patients ,including the annualized relapse rate (ARR),the Expanded Disability Status Scale(EDSS)and pregnancy outcomes. 70 NMOSD patients and 30 MS patients were screened out as information patients. The ARR and EDSS score in two groups were compared during the year before pregnancy,during pregnancy and after 1 year postpartum, respectively. 50 cases of normal pregnant women for the same period as the control group ,then to compared the difference of three groups on pregnancy outcomes.[Results]Attacks occurring during pregnancy or one year after childbirth/abortion in NMOSD and MS were 53.25%(41/77)and 20.00%(7/35)(P=0.001);The ARR during the first 3 months postpartum periods of NMOSD and MS group(2.65,2.51)was significantly higher than during the year before pregnancy(0.27,0.49,P < 0.001)and during pregnancy (0.32,0.2,P<0.001);The EDSS score of two groups increased after 1 year postpartum(3.06 ± 2.16,2.19 ± 1.28)than that during the year before pregnancy(1.58 ± 0.48,1.92 ± 1.29,P < 0.001)and during pregnancy(1.92 ± 1.35,1.67 ± 0.70,P < 0.001). There was no difference on ARR and EDSS score between NMOSD and MS group. NMOSD ,MS and normal control group had no dif?ference on pregnancy outcomes and neonatal weight.[Conclusions]Compared with MS,the attack of NMOSD had more closer relation?ship with pregnancy;both NMOSD and MS would increase the risk of disease relapsing and disability after pregnancy;the diseases had no effect on pregnancy outcomes.

Objective To explore the risk factors of postoperative activities of daily living in elderly patients after hip fracture. Methods By convenience sampling,from October 2013 to June 2015,150 elderly patients having undergone surgery after hip fracture were selected,with data of 141 cases finally collected. Gender,age,underlying diseases,surgical procedures,postoperative complications,ambulation time,whether walking with aids,family caregivers and reexamination conditions were recorded for evaluation of the patients about their activities of daily living half a year after the surgery. Mean comparison and chi-square test were used for univariate analysis about the above factors and Logistic regression analysis was used to identify possible risk factors. Results It was shown in univariate analysis that recovery of postoperative activities of daily living was influenced by age,surgical procedures,postoperative complications,ambulation time,walking aids,etc. (P< 0.05);it was shown in multivariate logistic regression that age (OR=1.065,95%CI:1.010-1.124, P=0.020),surgical procedures (OR=2.900,95%CI:1.200-7.008,P=0.018),postoperative complications (OR=6.366,95%CI:2.592-15.632,P< 0.001),ambulation time (OR=2.684,95%CI:1.133-6.359,P=0.025) and walking aids (OR=5.796,95%CI:1.542-21.780,P=0.009) were risk factors of postoperative daily living ability in elderly patients after hip fracture. Conclusions Old age,developing internal fixation surgery after incision,postoperative complications, walking more than 3 months after operation or walking without aids affect postoperative activities of daily living in elderly patients after hip fracture,thus affecting their quality of life, which makes it necessary for nursing staff to focus on risk factors for positive prevention and intervention.

Objective To investigate the change of serum nitric oxide ( NO ) in premature rupture of membrane (PROM) and its clinical significance, so as to assess the role of NO in the mechanism of PROM . Method The serum NO in 40 advanced stage pregnant women,28 premature delivery PROM,40 unknown reason term pregnancy PROM ,40 term pregnancy ,40 term labor were detected and compared. Results The serum NO was higher in unknown reason term pregnancy PROM [(95.364±3.562) (u)ol/L] than that in term pregnancy [(75.020±3.416)(u)ol/L] and term labor [(31.457±3.859)(u)ol/L](P<0.01). The serum NO was higher in premature delivery PROM [(101.256±3.124)(u)ol/L] than that in advanced stage pregnant women [(80.363±5.112)(u)ol/L] (P<0.0l),but similar with unknown reason term pregnancy PROM (P = 0.086). Conclusions The serum NO in PROM of term pregnancy significantly is higher than that of advanced stage and term labor women,indicates PROM relating to the increase of NO level. Because the increase of NO links to infection,it suggests that PROM relate closely to the amniotic infection.

Objective To explore the clinical characteristics,causes,diagnosis and treatment of pelvic mass in adolescents caused by gynecologic diseases. Methods One hundred and twenty-six adolescents with gynecologic diseases presented as pelvic mass were retrospectively reviewed. Results The average age of all patients was 17. 6 years,ranging from 10. 0 to 19. 0 years. Ectopic pregnancy was found in 38 patients(30. 2%) ,the average age of the patients was 18.6 ±0.5 years, ranging from 17.0 to 19.0 years. Ovarian neoplasm was found in 35 patients (27. 8%) ,the average age of the patients was 17. 1 ± 2. 1 years, ranging from 10. 0 to 19. 0 years. Twenty cases (57. l%)had germ cell tumor, eighteen with mature teratomas, one with endodermal sinus tumor and one with immature teratoma. Thirteen cases(37. 2%)had epithelial ovarian tumors;eight with serous cystadenomas,three with mucinous cystadenoma, one with mucinous cystadenoma complicated with serous cystadenoma and one with borderline mucinous cystadenoma. Two cases had simple cyst of the ovary (5. 7%) . Congenital abnormality of reproductive tract was found in 14 patients (11. 1%), the average age was 14. 7 ±2. 2 years, ranging from 13.0 to 19. 0 years. Seven had imperforate hymens, two had rudimentary horn of uterus,four had atresia of vagina,one had atresia of vagina complicated with rudimentary horn of uterus,and one had oblique septum of vagina. Corpus luteum rupture was found in 11 patients (8. 7%) , the average age was 18. 1 ± 1. 9 years, ranging from 13. 0 to 19. 0 years. 116 patients underwent operative treatment and 10 patients underwent conservative therapy. Conclusions According to the stage of adolescents, main causes of pelvic mass in adolescents are different Main causes of pelvic mass in early adolescents are tumorous disease. Congenital abnormalities of reproductive tract are usually in middle adolescents. The gonads develop closely mature in late adolescent period; main causes of pelvic mass are ectopic pregnancy, ovarian neoplasm and physiologic ovarian cyst Surgery is the main therapy of pelvic mass in adolescents.