[Objective] To explore the influencing factors of quality monitoring index on the nucleic acid pooling detection mode and continuously improve the detection quality of nucleic acid laboratory. [Methods] The quality monitoring indicators (NAT reactive rate, NAT resolution reactive rate, NAT invalid batch rate, NAT invalid result rate, equipment failure rate) and causes of invalidity in our laboratory from January 1, 2020 to December 31, 2022 were retrospectively analyzed. The quality monitoring indicators of the laboratory during 2020 to 2022 were compared longitudinally. The quality monitoring indicators of the laboratory in 2022 were compared horizontally with the overall level in Shandong for the same period to find the differences. [Results] From 2020 to 2022, a total of 218 686 samples were detected, the NAT reactive rate was 0.15‰ (32 samples in total), the resolution reactive rate was 39.02%, the invalid batch rate was 1.06%, the invalid result rate was 1.18%, and the equipment failure rate was 3.58%. There were no differences in the NAT reactive rate, NAT resolution reactive rate and NAT invalid batch rate among different years (P>0.05), but there were differences in the invalid result rate (P<0.05). Equipment failure was the main cause of invalid results (56.53%). Compared with other laboratories in Shandong, there were differences in the NAT reactive rate and invalid result rate (P<0.05). There were differences in the reaction rate, resolution rate and invalid result rate among different reagents (P<0.05). Compared with other two laboratories using the same manufacturer's reagent, there were differences in the reactive rate and invalid result rate (P<0.05), but no difference in the resolution rate and invalid batch rate (P>0.05). [Conclusion] Establishing quality indexes for process control and regular analysis can timely detect potential risks in laboratory operation. The use of quality indicators to implement self-comparison and inter-laboratory comparison can help the laboratory systematically and scientifically evaluate its own operating status and formulate corresponding quality management strategies, thereby improving the laboratory's testing capacity and ensure the safety of blood use.

Objective　To investigate the role of granulocyte-macrophage colony-stimulating factor (GM-CSF) deficiency in the pathogenesis of Exophiala oligosperma (E. oligosperma) infection, a dematiaceous fungus, aiming to provide new insights and evidence for the treatment of dematiaceous fungal infections. Methods　C57BL/6 wild-type (WT) mice and Csf2 gene knockout (KO) mice (C57BL/6 background) were selected. Using E. oligosperma isolated from patients with caspase recruitment domain-containing protein 9 (CARD9) gene deficiency, a murine subcutaneous infection model was established to simulate the human infection route. The natural progression of infection in the mice was observed for six weeks, with skin lesion tissues collected at appropriate time points for pathological analysis and monitoring immune responses. Results　Both WT and Csf2 KO mice exhibited spontaneous pathogen clearance and gradual recovery of foot tissue appearance during the progression of infection, with a 100% survival rate at the end of observation. Compared to WT mice, Csf2 KO mice showed reduced footpad swelling at 1 and 2 weeks post-infection (t=4.674, t=5.961, P<0.01). Fungal clearance in Csf2 KO mice was delayed, with fungal colonies still detectable in lesion tissues at week 4, and Periodic Acid-Schiff (PAS)-positive fungal spores observed in histopathological sections. There was no significant difference found in macrophage infiltration between WT and Csf2 KO mice during the early stages of infection (1-2 weeks) (P>0.05), while neutrophil infiltration was significantly reduced in Csf2 KO mice at week 2 (t=3.287, P<0.01). In addition, Csf2 KO mice exhibited lower levels of IL-6 and IL-1β in foot lesion homogenates at week 1 (t=4.686, t=4.102, P<0.05). Conclusions　This study demonstrated that GM-CSF deficiency delays pathogenic fungi clearance, prolongs the disease course and affects early inflammatory cytokine production as well as neutrophil infiltration during the early stages of fungal infections.

Objective:To investigate the effect of nuclear receptor subfamily 2 group F member 2(NR2F2)on the biological behaviors of human ovarian cancer SKOV3 cells,and to clarify its molecular mechauism and provide the new idea for treatment of ovarian cancer.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)Database analyse the expression level of NR2F2 gene in ovarian tissue,and analyse its correlation with clinical prognosis of ovarian cancer patients.The human ovarian cancer SKOV3 cells were divided into control group and NR2F2 over-expression(NR2F2 OE)group,which were transfected with mCherry control virus and NR2F2 OE over-expression virus,respectively,when the cell deusity reached 70％,and the stable transfection SKOV3 cell lines were screened with puromycin(puro)48h lafter.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the transfection efficiencies of the cells;RT-qPCR method was used to detect the expression levels of NR2F2 and sex-determining region Y-box 2(SOX2)mRNA in the cells in two groups;Western blotting method was used to detect the expression levels of NR2F2,ATP-binding cassette superfamily G member 2(ABCG2),and programmed cell death 1-ligand 1(PD-L1)protcins in the cells in two groups.CCK-8 assay was used to detect the proliferation activities of the cells in two groups;Wound assay was used to detect the migration rates of the cells in two groups;Transwell chamber assay was used to detect the number of transmembrane cells;Spheroidization assay was used to detect the numbers of spheroids in the cells;peripheral blood mononuclear cells(PBMCs)-mediated tumor cell killing assay was used to detect the relative densities of surviving tumor cells;CCK-8 assay was used to detect the half maximal inhibitory concentration(IC50)of paclitaxel(PTX)and carboplatin(CBP).Results:Compared with normal ovarian tissue,the expression level of NR2F2 gene in ovarian tumor tissue was decreased(P＜0.05),and decreased with the improvement of clinical pathological grading of ovarian tumor.The patients with higher expression level of NR2F2 gene had better clincal prognosis.The SKOV3 cells with NR2F2 over-expresson were successfully constructed,and the expression levels of NR2F2 mRNA and protein in the cells in NR2F2 OE group were increased compared with control group(P＜0.001).The CCK-8 assay results showed that compared with control group,the proliferation activities of the cells in NR2F2 OE group were decreased at different time points(1,2,3,and 4 d)(P＜0.05 or P＜0.01).The cell wound assay results showed that compared with control group,the migration rate of the cells in NR2F2 OE group was decreased(P＜0.001).The Transwell assay results showed that compared with control group,the number of transmembrane cells in NR2F2 OE group was decreased(P＜0.01).Compared with control group,the number of the spheroids in NR2F2 OE group was decreased(P＜0.05),and the expression levels of SOX2 mRNA(P＜0.01)and protein(P＜0.001)were increased.Compared with control group,the relative density of surviving tumor cells in NR2F2 OE group was decreased,but the difference was not significant(P＜0.05),and the expression level of PD-L1 protein was decreased(P＜0.05).Compared with control group,the proliferation activities of cells in NR2F2 OE group were decreased(P＜0.05),and the drug sensitivities of the cells to PTX and CBP were enhanced(P＜0.05);the IC50 of PTX was significantly reduced,while the IC50of CBP could not be calculated due to excessively high drug concentration;the expression level of ABCG2 protein was decreased(P＜0.05).Conclusion:The over-expression of NR2F2 may inhibit the proliferation,migration,and invasion of the human ovarian cancer SKOV3 cells,decrease the expression levels of SOX2,PD-L1 and ABCG2 proteins,suppress the stemness and immune evasion ability of the SKOV3 cells,and enhance the sensitivities of the SKOV3 cells to PTX and CBP.

Cerebral blood flow directly affects the metabolism of substances and neural activity in the brain, and is closely associated with the occurrence and development of cerebral small vessel disease (CSVD). Multiple studies have revealed that various imaging biomarkers in patients with CSVD, such as lacunar infarction, enlarged perivascular spaces, cerebral microbleeds, cerebral atrophy, and white matter hyperintensities, are closely associated with cerebral autoregulation (CA) function. Therefore, understanding the regulatory mechanism of CA in patients with CSVD is of great significance for delaying the further development of CSVD, improving cerebral ischemia and cognitive impairment. This article reviews the correlation and mechanism between CA and CSVD.

Objective:To investigate the effect of abdominal circumference on intestinal radiation dose volume and acute intestinal toxicity in pelvic intensity modulated radiation therapy for rectal cancer.Methods:A total of 150 patients with locally advanced rectal cancer (LARC) who received adjuvant and neoadjuvant concurrent chemoradiotherapy at the Affiliated Cancer Hospital of Guizhou Medical University from March 2023 to January 2025 were enrolled, including 82 cases of adjuvant radiotherapy and 68 cases of neoadjuvant radiotherapy. All patients underwent radiotherapy CT simulation positioning in the standard mode of prone position with abdominal board padding and bladder filling. Intestinal toxicity was categorized as a binary variable based on the occurrence of ≥2 grade acute intestinal toxicity. Linear and logistic regression models were used to analyze the factors influencing intestinal radiation dose volumes (V 10, V 20, V 30, V 40) and acute intestinal toxicity in LARC patients. Generalized additive models and piecewise linear and logistic regression analyses were employed to examine the threshold effects of abdominal circumference on intestinal radiation dose volumes and acute intestinal toxicity. The threshold value for abdominal circumference was determined based on the upper limit of the 95% CI for the threshold. A difference test was used to validate the differences in intestinal radiation dose volume and acute intestinal toxicity between small and medium-to-large abdominal circumferences. Results:Univariate analysis showed that, gender, body mass, abdominal circumference, planning target volume (PTV), intestinal volume were all influencing factors for the radiation dose volumes (V 10, V 20, V 30, V 40) of each intestinal segment of patients with LARC undergoing adjuvant radiotherapy (all P<0.05). Body mass, abdominal circumference, intestinal volume were all influencing factors for the radiation dose volumes (V 10, V 20, V 30, V 40) of each intestinal segment of patients with LARC undergoing neoadjuvant radiotherapy (all P<0.05). Body mass index (BMI), abdominal circumference, intestinal volume and individual intestinal radiation volumes (V 10, V 20, V 30, V 40) were all influencing factors for the acute intestinal toxicity of patients with LARC undergoing adjuvant radiotherapy (all P<0.05). Body mass, BMI, abdominal circumference, multiple intestinal radiation dose volumes (V 20, V 30, V 40) were all influencing factors for the acute intestinal toxicity of patients with LARC undergoing neoadjuvant radiotherapy (all P<0.05). Multivariate analysis showed that, abdominal circumference (V 10: β=-1.01, 95% CI: -1.68--0.33, P=0.004; V 20: β=-0.94, 95% CI: -1.28--0.60, P<0.001; V 30: β=-0.58, 95% CI: -0.82--0.34, P<0.001; V 40: β=-0.41, 95% CI: -0.60--0.23, P<0.001) was an independent influencing factor for the radiation dose volume of each intestinal segment of patients with LARC undergoing adjuvant radiotherapy. Abdominal circumference (V 10: β=-0.92, 95% CI: -1.62--0.22, P=0.010; V 20: β=-0.84, 95% CI: -1.11--0.57, P<0.001; V 30: β=-0.42, 95% CI: -0.57--0.28, P<0.001; V 40: β=-0.30, 95% CI: -0.41--0.19, P<0.001) was an independent influencing factor for the radiation dose volume of each intestinal segment of patients with LARC undergoing neoadjuvant radiotherapy. Abdominal circumference ( OR=0.86, 95% CI: 0.78-0.95, P=0.002) was an independent influencing factor for the acute intestinal toxicity of patients with LARC undergoing adjuvant radiotherapy. Abdominal circumference ( OR=0.87, 95% CI: 0.79-0.96, P=0.004) was an independent influencing factor for the acute intestinal toxicity of patients with LARC undergoing neoadjuvant radiotherapy. The generalized additive model revealed a nonlinear relationship between abdominal circumference and intestinal radiation dose volume and acute intestinal toxicity of adjuvant radiotherapy patients. Further segmented regression analysis results showed that there was a threshold effect between abdominal circumference and intestinal radiation dose volume (V 10, V 20, V 30, V 40) and acute intestinal toxicity. The inflection point values between abdominal circumference and intestinal radiation dose volume V 10, V 20, V 30, V 40 in LARC patients undergoing adjuvant radiotherapy were all 71.9 cm; the inflection point values between abdominal circumference and the intestinal radiation dose volume V 10, V 20, V 30, V 40 in LARC patients undergoing neoadjuvant radiotherapy were 69.0, 69.0, 69.0, 68.6 cm, respectively; The inflection point values between abdominal circumference and acute intestinal toxicity in LARC patients undergoing adjuvant radiotherapy and neoadjuvant radiotherapy were 71.9, 69.0 cm, respectively. Based on the upper limit of the 95% CI threshold, the cutoff values for small and medium-to-large abdominal circumferences for patients undergoing adjuvant and neoadjuvant radiotherapy were set at 76.1, 71.9 cm, respectively. In patients undergoing adjuvant radiotherapy, the levels of intestinal radiation dose volume V 10 [ (7.65±2.29) cm 3vs. (5.88±2.68) cm 3, t=2.76, P=0.007], V 20 [ (4.28±1.27) cm 3vs. (2.72±1.31) cm 3, t=4.81, P<0.001], V 30 [ (2.42±1.07) cm 3vs. (1.37±0.76) cm 3, t=4.95, P<0.001], V 40 [ (1.69±0.74) cm 3vs. (0.92±0.58) cm 3, t=4.93, P<0.001] in the small abdominal circumference group ( n=22) were significantly higher than those in patients with medium-to-large abdominal circumferences ( n=60) ; In patients undergoing neoadjuvant radiotherapy, patients with small abdominal circumferences ( n=11) had significantly higher V 20 [ (3.09±0.84) cm 3vs. (2.28±1.17) cm 3, t=2.17, P=0.033], V 30 [1.44 (1.22, 1.53) cm 3vs. 0.91 (0.56, 1.22) cm 3, Z=-3.04, P=0.002], V 40 [0.93 (0.84, 1.09) cm 3vs. 0.44 (0.30, 0.81) cm 3, Z=-3.19, P=0.001] than patients with medium-to-large abdominal circumferences ( n=57). In patients receiving adjuvant radiotherapy and neoadjuvant radiotherapy, there were statistically significant differences in acute intestinal toxicity between patients with small abdominal circumferences and with medium-to-large abdominal circumferences ( χ2=10.46, P=0.001; χ2=8.13, P=0.004) . Conclusions:In the standard mode (prone position with abdominal board padding and bladder filling), abdominal circumference is an independent factor influencing the intestinal radiation dose volume and acute intestinal toxicity in rectal cancer radiotherapy patients. There is a significant non-linear threshold effect between abdominal circumference and different levels of intestinal radiation dose volume and acute intestinal toxicity. The impact of abdominal circumference on intestinal radiation dose volume and toxicity differs significantly before and after the inflection point value. Patients with smaller abdominal circumferences not only fail to achieve the expected benefits under the current standard radiotherapy regimen but also face higher risks of intestinal radiation dose volume and toxicity.

Objective To find out whether photobiomodulation(PBM)can mitigate cognitive dysfunction caused by chronic stress by affecting levels of adenosine triphosphate(ATP)and adenosine receptors.Methods Twenty-four C57BL/6J mice were randomly divided into a control group,a stress group,and a treatment group.Chronic unpredictable mild stress was used to establish a mouse model of stress.Six weeks into modeling,the treatment group was subjected to one week of PBM interventions.Behavioral tests were conducted to observe behavioral changes in the mice.Western blotting(WB)was used to detect the expressions of A1,A2B,and A3 adenosine receptors in the hippocampus and prefrontal cortex of mice in the three groups.Twelve C57BL/6J mice were randomly divided into a control group and an intervention group.The intervention group received a week of PBM interventions and underwent behavioral testing.WB was used to detect the expression changes of A1,A2B,and A3 adenosine receptors in the hippocampus and prefrontal cortex in both groups.Immunofluorescence assay was adopted to detect the expression of c-Fos in the hippocampus of mice in the two groups.The ATP assay kit made by Beyotime Biotechnology Co.,Ltd.was used to measure changes in ATP contents in the hippocampus and prefrontal cortex tissues of mice.Cell experiments were conducted to verify the effect of PBM on intracellular ATP contents.Results Mice in the stress group covered a similar distance to the control group,but finished far fewer platform crossings.There was no significant difference between the treatment group and the control group in the number of times of platform crossings,but compared favorably with the stress group where the levels of adenosine receptors in the hippocampus and prefrontal cortex were lower,but were increased by PBM.After PBM interventions in normal mice,platform crossings were increased significantly compared to the control group.PBM also raised adenosine receptor levels and ATP contents in the hippocampus and prefrontal cortex,and increased hippocampal c-Fos expressions.In vitro,PBM elevated intracellular ATP levels.Conclusion PBM may improve chronic stress-induced cognitive dysfunction by regulating ATP levels and adenosine receptor expressions,thereby modulating neuronal responsiveness in the hippocampus.

Objective To investigate the survival time of patients diagnosed with sporadic Creutzfeldt-Jakob disease in China between 2020 and 2022 and explore the associated factors influencing survival.Methods A retrospective analysis was conducted on clinically diagnosed cases with complete information on sporadic Creutzfeldt-Jakob dis-ease diagnosed by the China Creutzfeldt-Jakob disease surveillance network from 2020 to 2022,baseline information of patients was obtained from the case files,telephone follow-up was used to obtain the treatment and survival status of the patients after the diagnosis,life-table method was used for estimating the survival rate,Kaplan-Meier method was used for calculating the median survival time and the 95％CI,Cox regression model was used for univariate and multivariate analyses were used to screen for factors influencing survival time.Results The median survival time of the 300 patients was 5 months(95％CI:4.165-5.835).Univariate analysis revealed that factors such as age at onset,regional distribution,presence of corticobasal or extrapyramidal symptoms as initial manifestations,number of initial symptoms,presence of corticobasal or extrapyramidal functional abnormalities,number of major clinical manifestations,presence of typical electroencephalogram findings,and use of nasal feeding during the course of the disease were potential factors influencing survival time(P＜0.1).Multivariate analysis showed that the risk of death in patients with onset age＞65 years was 1.350 times higher than in patients with onset age ≤65 years(P=0.021,95.0％CI:1.046-1.742).Patients without pyramidal or extrapyramidal dysfunction had a 0.674-fold lower risk of death compared to those with these symptoms(P=0.020,95.0％CI:0.483-0.939).Patients who did not receive nasal feeding had a 1.817-fold higher risk of death compared to those who did(P＜0.001,95.0％CI:1.406-2.349).Conclusion Age at onset,the presence of pyramidal or extrapyramidal functional abnormalities,and the use of nasal feeding during the disease course are factors influencing the survival time of pa-tients clinically diagnosed with sCJD.

Objective:To understand the medical care of patients with sporadic Creutzfeldt-Jakob disease in China and its relationship with survival time.Methods:A retrospective analysis was performed on data of 150 patients with sporadic Creutzfeldt-Jakob disease diagnosed by China′s Creutzfeldt-Jakob Disease Surveillance Network during the period of January 1, 2021 to December 31, 2022 in this study, and telephone follow-up with family members was used to obtain information of the patients′ care, treatment, and survival after diagnosis. Survival was estimated by life table method, median survival time and 95% confidence interval ( CI) were calculated by Kaplan-Meier method, log-rank method was used to compare the difference in survival time between different groups, and multifactorial analysis was performed by COX proportional risk regression model regarding the influencing factors on patients′ survival time. Results:The median survival time of 150 patients with sporadic Creutzfeldt-Jakob disease was 6 months, and the cumulative lifetime survival rates at 2, 6, 12, and 18 months were 62%, 39%, 22%, and 9%, respectively. The result of univariate analysis showed that the differences in survival time between groups with the presence or absence of cortical blindness in the first symptom, the presence or absence of respiratory support (oxygen therapy), the presence or absence of adjunctive medication, and the presence or absence of tube feeding (nasogastric) were meaningful ( P<0.1). Multifactorial COX regression analysis showed that the risk of death in patients without adjuvant medication was 1.429 times higher than that in patients with adjuvant medication (95.0% CI: 1.014-2.014), and the risk of death in patients without tube feeding (nasal feeding) was 1.479 times higher than that in patients with tube feeding (nasal feeding) (95% CI: 1.052-2.081). Conclusions:Whether or not adjuvant medication is administered and whether or not tube feeding (nasogastric) is used are factors that affect survival time in patients with sporadic Creutzfeldt-Jakob disease, and the administration of appropriate adjuvant medication and tube feeding (nasogastric) may contribute to prolonging survival time in patients with sporadic Creutzfeldt-Jakob disease.

Objective To compare the value of TACE+microwave ablation(MWA)+programmed death-1(PD-1)and TACE+MWA for treating primary hepatic carcinoma(PHC).Methods Data of 80 PHC patients who underwent TACE+MWA+PD-1(observe group)or TACE+MWA treatment(control group)were retrospectively analyzed(each n=40).The baseline data,therapeutic efficacy of PHC and complications were compared between groups.Results No significant difference of baseline data was found between groups(all P＞0.05).The objective response rate(ORR)of observe group and control group was 90.00％and 72.50％,respectively,while the disease control rate(DCR)was 97.50％and 95.00％,respectively,with no significant difference between groups(both P＞0.05).The median overall survival(OS)of observe group and control group was 30.80 months and 15.70 months,respectively,while the median progression free survival(PFS)was 23.35 months and 6.80 months,respectively.OS and PFS of observe group were both longer than those of control group(both P＜0.05).No significant difference of the incidence of complications was detected between groups(all P＞0.05),and all were improved after symptomatic treatment.Conclusion TACE+MWA+PD-1 was superior to TACE+MWA for treating PHC.

Objective To explore the relationship between the expression of Toll like receptor 4(TLR4)and Janus protein tyrosine kinase 3(JAK3)genes and the imbalance of helper T cell 17(Th17)/regulatory T cells(Treg)in peripheral blood mononuclear cells(PBMCs)of patients with ankylosing spondylitis(AS).Methods A total of 101 AS patients were selected as the AS group.According to the AS Disease Activity Index(BASDAI)score,patients were divided into the AS stable phase(ASS)group(＜4 points,42 cases)and the AS active phase(ASA)group(≥4 points,59 cases).Additionally,50 healthy volunteers who underwent physical examinations at our hospital during the same period were selected as the control group(health group).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to determine the mRNA expression of TLR4 and JAK3 in PBMCs,and the proportion of serum Th17 and Treg was measured and Th17/Treg levels were calculated.Results The levels of TLR4 mRNA and JAK3 mRNA in PBMCs of peripheral blood were higher in the AS group than those in the health group(P＜0.05).The levels of Th17,proportion and Th17/Treg in peripheral blood were higher in the AS group than those of the health group(P＜0.05).Higher levels of TLR4 mRNA,JAK3 mRNA,Th17,and Th17/Treg were risk factors for AS,while higher level of Treg was protective factors for AS.TLR4 mRNA and JAK3 mRNA in AS patients were positively correlated with Th17 ratio and Th17/Treg ratio,and negatively correlated with Treg ratio(P＜0.05).Conclusion TLR4 and JAK3 genes are highly expressed and Th17/Treg is unbalanced in PBMC of AS patients.The high expression of TLR4 and JAK3 genes is closely related to Th17/Treg imbalance in AS patients.