Objective To investigate the impact of Toxoplasma gondii type I, II and III rhoptry protein 16 (ROP16) on programmed cell death ligand 1 (PD-L1) expression in lung adenocarcinoma cells, and to examine the effects of T. gondii type I ROP16 protein on the relative PD-L1 expression, the relative PD-L1 distribution on the cell membrane surface, and the binding of programmed cell death 1 (PD-1) to PD-L1 in lung adenocarcinoma cells. Methods Lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins were generated, and transfected into the human lung adenocarcinoma A549 cell line. A549 cells were used as a blank control group, and A549 cells transfected with an empty lentiviral expression vector were used as a negative control group, while A549 cells transfected with lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins served as experimental groups. Stably transfected cells were selected with puromycin and verified using Western blotting, quantitative real-time PCR (RT-qPCR), and immunofluorescence assays. The PD-L1 expression was quantified at translational and transcriptional levels using Western blotting and RT-qPCR assays in A549 cells in the five groups, and the relative PD-L1 distribution was detected on the A549 cell membrane surface using flow cytometry. In addition, the effect of T. gondii type I ROP16 protein on the PD-1/PD-L1 binding was measured in A549 cells using enzyme-linked immunosorbent assay (ELISA). Results The relative ROP16 protein expression was 0, 0, 1.546 ± 0.091, 1.822 ± 0.047 and 2.334 ± 0.089 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 1 339.00,P < 0.001), and the relative ROP16 mRNA expression was 2.153 ± 0.949, 2.436 ± 1.614, 14.343 ± 0.020, 12.577 ± 0.285 and 15.090 ± 0.420 in the blank control group, negative control group and the T. gondii type I, II and III ROP16 protein overexpression groups, respectively (F = 483.50,P < 0.001). The ROP16 expression was higher in the T. gondii type I, II and III ROP16 protein overexpression groups than in the blank control group at both translational and transcriptional levels (allP values < 0.001). Immunofluorescence assay revealed that T. gondii type I, II and III ROP16 proteins were predominantly localized in A549 cell nuclei. Western blotting showed that the relative PD-L1 protein expression was 0.685 ± 0.109, 0.589 ± 0.114, 1.007 ± 0.117, 0.572 ± 0.151, and 0.426 ± 0.116 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 9.46,P < 0.05), and RT-qPCR assay quantified that the relative PD-L1 mRNA expression was 1.012 ± 0.190, 1.281 ± 0.465, 1.950 ± 0.175, 0.889 ± 0.251, and 0.230 ± 0.192 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 14.18,P < 0.05). The PD-L1 expression was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group at both translational and transcriptional levels (both P values < 0.05). Flow cytometry detected that the relative distributions of PD-L1 protein were (10.83 ± 0.60)%, (11.23 ± 0.20)%, and (14.61 ± 0.50)% on the A549 cell membrane surface (F = 28.31, P < 0.05), and the relative distribution of PD-L1 protein was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group and negative control group (both P values < 0.001). ELISA measured significant differences in the absorbance (A) value among the T. gondii type IROP16 protein overexpression group, the blank control group and the negative control group if the concentrations of the recombinant PD-1 protein were 0.04 (F = 10.45, P < 0.05), 0.08 μg/mL (F = 11.68, P < 0.05) and 0.12 μg/mL (F = 52.68, P < 0.05), and the A value was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group and the negative control group (both P values < 0.05), indicating that T. gondii type IROP16 protein promoted the PD-L1/PD-1 binding in A549 cells in a concentration-dose manner. Conclusions T. gondii type IROP16 protein overexpression may up-regulate PD-L1 expression in A549 cells at both transcriptional and translational levels and the relative PD-L1 distribution on the A549 cell membrane surface, and affect the PD-1/PD-L1 binding in a concentration-dependent manner.

Background Photovoltaic (PV) cells can convert solar energy into electricity and alleviate the dilemma of energy supply shortage. Shanghai's PV module industry is booming, but there are few reports on the health status of the workers and there is an urgent need for health risk assessment. Objective To evaluate the health status of PV module production workers and the association between work and health status, in order to provide a direction for the health management of the workers. Methods Among the PV module production workers who completed prescriptive occupational health examination by a designated medical institution in 2021, 2453 workers with more than one year of working age were selected as the exposure group and 538 newly hired workers as the control group. On the basis of the Technical specifications for occupational health surveillance (GBZ 188−2014), the health examination included physical examination and laboratory examination and information such as sociodemographics, living habits, and disease history. We compared the indicators of pure-tone hearing test, blood routine examination, electrocardiogram (ECG), liver function, and kidney function between the two groups. The blood routine results included erythrocyte-related indicators, leukocyte-related indicators, and platelet-related indicators, and the results of liver function included hepatocyte injury indicators, hepatic secretory function indicators, and hepatic synthesis function indicators. The workers were divided into four groups by quartiles of working age. Trend chi-square test was used to analyze differences in health status between the four working age groups. Multiple logistic regression models were used to evaluate the association between working age and health indicators. Results Among the exposure group workers, 2035 (83.0%) were male and 418 (17.0%) were female. The median (P₂₅, P₇₅) age was 34.0 (30.0, 39.0) years and the median (P₂₅, P₇₅) working age was 6.0 (3.0, 10.0) years. The abnormality rate of blood routine was 61.7%. Among them, the abnormality rates of platelet-related indicators and liver secretory function indicators were 39.8% and 48.1% respectively. The risks of abnormal hepatocyte injury-related indicators, fatty liver, and platelet-related indicator abnormalities among the exposure group were 1.471 (95%CI: 1.060, 2.054), 1.691 (95%CI: 1.208, 2.385), and 7.576 (95%CI: 4.967, 11.994) times higher than those in the control group respectively. The single-factor analysis demonstrated a positively linear trend between working age and hypertension prevalence, electrical audiometry abnormality rate, or liver secretory function indicator abnormality rate. Corrected for gender, age, smoking status, hypertension, etc., the results of logistic analysis showed that quartile working age was positively related to abnormal liver secretion function and abnormal platelet-related indicators respectively (OR=1.047, P=0.005; OR=1.037, P=0.014), and inversely associated with the abnormal rate of renal function (OR=0.953, P=0.044). Conclusion Negative associations between health status and working age are identified in PV module production workers. The target PV module production employees are in younger age, and with the increase of working age, the abnormalities of liver function and platelets may increase. Therefore, the enterprises should extend occupational health work from workplace to workers.

Background::There is a need for effective and safe therapies for psoriasis that provide sustained benefits. The aim of this study was to assess the efficacy and safety of tildrakizumab, an anti-interleukin-23p19 monoclonal antibody, for treating moderate-to-severe plaque psoriasis in Chinese patients.Methods::In this multi-center, double-blind, phase III trial, patients with moderate-to-severe plaque psoriasis were enrolled and randomly assigned (1:1) to receive subcutaneous tildrakizumab 100 mg or placebo at weeks 0 and 4. Patients initially assigned to placebo were switched to receive tildrakizumab at weeks 12, 16, and every 12 weeks thereafter. Patients in the tildrakizumab group continued with tildrakizumab at week 16, and every 12 weeks until week 52. The primary endpoint was the Psoriasis Area and Severity Index (PASI 75) response rate at week 12.Results::At week 12, tildrakizumab demonstrated significantly higher PASI 75 response rates (66.4% [73/110] vs. 12.7% [14/110]; difference, 51.4% [95% confidence interval (CI), 40.72, 62.13]; P <0.001) and Physician’s Global Assessment (60.9% [67/110] vs. 10.0% [11/110]; difference, 49.1% [95% CI, 38.64, 59.62]; P <0.001) compared to placebo. PASI 75 response continued to improve over time in both tildrakizumab and placebo-switching to tildrakizumab groups, reaching maximal efficacy after 28 weeks (86.8% [92/106] vs. 82.4% [89/108]) and maintained up to 52 weeks (91.3% [95/104] vs. 87.4% [90/103]). Most treatment-emergent adverse events were mild and not related to tildrakizumab. Conclusion::Tildrakizumab demonstrated durable efficacy through week 52 and was well tolerated in Chinese patients with moderate-to-severe plaque psoriasis.Trial registration::ClinicalTrials.gov, NCT05108766.

N1-methyladenosine(m1A)RNA methylation is critical for regulating mRNA translation;however,its role in the development,progression,and immunotherapy response of head and neck squamous cell carcinoma(HNSCC)remains largely unknown.Using Tgfbr1 and Pten conditional knockout(2cKO)mice,we found the neoplastic transformation of oral mucosa was accompanied by increased m1A modification levels.Analysis of m1A-associated genes identified TRMT61A as a key m1A writer linked to cancer progression and poor prognosis.Mechanistically,TRMT61A-mediated tRNA-m1A modification promotes MYC protein synthesis,upregulating programmed death-ligand 1(PD-L1)expression.Moreover,m1A modification levels were also elevated in tumors treated with oncolytic herpes simplex virus(oHSV),contributing to reactive PD-L1 upregulation.Therapeutic m1A inhibition sustained oHSV-induced antitumor immunity and reduced tumor growth,representing a promising strategy to alleviate resistance.These findings indicate that m1A inhibition can prevent immune escape after oHSV therapy by reducing PD-L1 expression,providing a mutually reinforcing combination immunotherapy approach.

AIM:To investigate the effects of Xiaozhong-Zhitong mixture(XZZT)on M2 polarization and Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway in mouse microglia(BV2 cells).METHODS:The BV2 cells were divided into 5 groups:blank group,model group[lipo-polysaccharide(LPS)+hypoxia],TAK-242(resatorvid,a TLR4 inhibitor)group(LPS+hypoxia+TAK-242),XZZT group(LPS+hypoxia+XZZT),and TAK-242+XZZT group(LPS+hypoxia+TAK-242+XZZT).Flow cytometry was used to detect early apoptosis and cell cycle of BV2 cells,and immunofluorescence staining was employed to detect the positive expres-sion of M1-type marker inducible nitric oxide synthase(iNOS)and M2-type marker CD206.Western blot was utilized to detect the expression of TLR4/MyD88/NF-κB signaling pathway-related proteins,including TLR4,MyD88,NF-κB p65,phosphorylated p65(p-p65),phosphorylated transforming growth factor-β-activated kinase 1(p-TAK1),and phosphory-lated IκB kinase α/β(p-IKKα/β).RT-qPCR was used to detect the mRNA expression of interleukin-1β(IL-1β),IL-10,tumor necrosis factor-α(TNF-α),TLR4,MyD88,and NF-κB p65.RESULTS:Compared with model group,the rate of early apoptosis was significantly decreased in XZZT group(P<0.01),the percentage of cells arrested in the S phase was significantly increased(P<0.01),and the protein levels of TLR4,MyD88,NF-κB p65,p-IKKα/β,p-p65,and p-TAK1 were significantly decreased(P<0.05 or P<0.01).Additionally,IL-1β,TNF-α,TLR4,MyD88 and NF-κB p65 mRNA expression levels were significantly decreased(P<0.05 or P<0.01),while IL-10 mRNA expression was significantly in-creased(P<0.05).Compared with TAK-242 group,the average percentage of iNOS positive area was significantly de-creased,while CD206 was significantly increased in TAK-242+XZZT group(P<0.01).CONCLUSION:The XZZT has the effect of inducing M2 polarization of mouse microglia,and the mechanism may be linked to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

Objective:To investigate the facilitators and hindrances of smoking cessation in patients with chronic obstructive pulmonary disease (COPD), and to provide a basis for developing individualized smoking cessation intervention strategies for COPD patients.Methods:Based on the health ecology theory and using a phenomenological approach in qualitative research, purposive sampling was used to select 15 COPD patients with smoking history who were admitted to Tianjin Fourth Central Hospital from March to May 2023 and face-to-face semi-structured interviews were conducted. Colaizzi seven-step analysis was used to analyze the interview content.Results:A total of 15 COPD patients were interviewed, including 13 males and 2 females, aged 61-75 years old. The facilitators of smoking cessation in COPD patients included complications, sequelae of novel coronavirus infection, fear of death, smoking cessation counseling by medical staff, heavy family financial burden, and smoke-free environment. The hindrances of smoking cessation in COPD patients included milder disease symptoms of COPD, higher levels of nicotine dependence, false disease-related perceptions, family supervision and control, and occupational experience.Conclusions:Smoking cessation in COPD patients is influenced by five factors: personal characteristics, behavioral characteristics, interpersonal network, living and working conditions, and policy environment. Medical staff and relevant national institutions should formulate corresponding smoking cessation strategies according to address the facilitators and hindrances of smoking cessation in COPD patients, so as to further reduce the smoking prevalence of COPD patients, promote the health of patients and reduce the burden of disease.

Background Flurochloridone (FLC) is toxic to male reproduction and can induce apoptosis of testicular tissue and supporting cells under oxidative stress. Of particular concern is whether nuclear factor-erythrocyte 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway and nuclear factor kappa B (NFκB) signaling pathway participate this process. Objective To observe apoptosis of testicular tissue and sertoli TM4 cells and alterations of Nrf2/HO-1 and NFκB signaling pathways in mice treated with FLC in vivo/in vitro. Methods (1) Animal experiment. Testis samples were harvested from male C57BL/6 mice after 28-day FLC (0, 3, 15, 75, and 375 mg·kg⁻¹ per day) exposure via oral route. Malondialdehyde (MDA) and superoxide dismutase (SOD) in homogenate of testicular tissue were measured by colorimetry. Apoptosis of testicular tissue was evaluated by TUNEL staining. Expression and distribution of Nrf2 and NFκB were detected by immunohistochemistry. Protein expression levels of Nrf2, HO-1, NAD(P)H: quinone oxidoreductase 1 (NQO1), NFκB, inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ), and phosphorylated recombinant inhibitory subunit of nuclear factor kappa-B alpha (P-IκBα) in testicular tissue homogenate were determined by Western blotting. (2) Cell experiment. TM4 cell lines were treated with 40, 80, 120, 160, and 200 μmol·L⁻¹ FLC for 6 h, and cell viability was detected by CCK-8. After 6 h exposure to 40, 80, and 160 μmol·L⁻¹ FLC, the apoptosis rate was detected by flow cytometry, and the protein expression levels of Nrf2, HO-1, NQO1, NFκB, IKKβ, and IκBα were detected by Western blotting. Results (1) Animal experiment. Apoptosis occurred in the interstitial and basal parts of spermatogenic tubules in male C57BL/6 mice after 28 days of oral FLC exposure. Compared with the control group, the MDA level in testicular tissue of the 375 mg·kg⁻¹ FLC-treated group was significantly increased (P<0.05), and the SOD activity was significantly decreased (P<0.05). After 375 mg·kg⁻¹ FLC exposure, apoptosis occurred in the interstitial and basal parts of spermatogenic tubules. The results of immunohistochemistry showed the expression of Nrf2 and NFκB in the interstitium and basal part of spermatogenic tubules of the treated groups. Compared with the control group, the protein levels of Nrf2, NQO1, P-IκBα, NFκB, and IKKβ in the 15, 75, and 375 mg·kg^-1 groups were significantly increased (P<0.001), and the HO-1 protein level was significantly increased in the 375 mg·kg⁻¹ group (P<0.001). (2) Cell experiment. Compared with the control group, the TM4 cell viabilities in the 40, 80, 120, 160, and 200 μmol·L⁻¹ FLC-treated groups significantly decreased (P<0.01). The apoptosis rates were significantly increased (P<0.05), and the apoptosis rates increased from 5.7% in the control group to 7.4%, 9.4%, and 11.7% in the 40, 80, and 160 μmol·L⁻¹, respectively. The Nrf2 protein level in the 40 μmol·L⁻¹ group was significantly increased (P<0.01), while the levels significantly decreased in the 80 and 160 μmol·L⁻¹ groups (P<0.01). The HO-1 protein levels in the 40, 80, and 160 μmol·L⁻¹ groups were significantly increased (P<0.01). The level of NQO1 protein in the 40 μmol·L⁻¹ group was significantly increased (P<0.01). The NFκB protein levels were significantly increased in the 80 and 160 μmol·L⁻¹ groups (P<0.001). The IκBα protein levels were significantly decreased in all treated groups (P<0.001). The IKKβ protein had no significant change. Conclusion FLC induces testicular tissue apoptosis, and the process affects Nrf2/HO-1 signaling pathway and NFκB signaling pathway. The in vitro study confirms that FLC could induce apoptosis of TM4 cells and activate Nrf2/HO-1 and NFκB signaling pathways.

Homozygous familial hypercholesterolemia (HoFH) is a rare and serious autosomal genetic metabolic disease. Patients without intervention often die younger than 30 years old from early atherosclerotic cardiovascular disease (ASCVD)incurred by extremely high levels of low-density lipoprotein cholesterol (LDL-C). We present a case of HoFH, a child with compound heterozygous mutation in this study. The effect of conventional lipid-lowering therapy through diet control and lipid-lowering drugs was unsatisfactory. The blood-lipid purification proves effective but has poor compliance and difficult to maintain for a longer time. The patient received orthotopic liver transplantation and had been followed for 2 years, with the patient shows normal LDL-C, well growth and development. We hope the case will provide the clinician with better understanding of the diagnosis and treatment of the rare disease of HoFH.

Purpose To evaluate the fatty infiltration of lower limbs by using iterative decomposition of water and fat with asymmetry and least squares estimation quantitative fat imaging(IDEAL-IQ)technique in idiopathic inflammatory myopathy(IIM)patients,and to analyze the correlation between muscle fat fraction(FF)and clinical assessments.Materials and Methods Thirty-two patients with IIM were diagnosed by muscle biopsy and 32 age-,gender-matched healthy volunteers(control group)were recruited.T1WI,T2WI in axial view and IDEAL-IQ sequence of thighs were scanned on each subject.FF values of the anterior,interior and posterior thigh muscles were measured on the FF image generated in the IDEAL-IQ sequence,and medical research council scale score of the IIM group were collected.The difference of muscle FF value between the IIM group and control group was compared,the correlation between FF value and muscle strength of thigh muscles was also analyzed.Results The mean FF values of anterior,interior and posterior thigh muscles in IIM group were 16.60±3.67,6.77±4.92 and 17.32±4.01,respectively,which were significantly higher than those in control group(2.58±2.57,1.40±0.64 and 1.57±0.19),with statistically significant differences(t=-7.29,-6.91,-4.85;all P<0.05).Spearman test showed that the mean FF value was significantly correlated with course of disease(r=0.587,P<0.001).The mean FF values of anterior,interior and posterior thigh muscles were significantly correlated with muscle strength(r=-0.885,-0.761,-0.594;all P<0.001).Conclusion The IDEAL-IQ technique can quantitatively and objectively analyze the severity of muscle fat infiltration in IIM patients,and its degree is correlated with the muscle strength,which has significant clinical application value.

Objective:To investigate the cosmetic effects of botulinum toxin type A(BTX-A) in the prevention and treatment of cicatrix on split-thickness skin graft donor site.Methods:A prospective randomized controlled study was commenced to recruit patients with functional burns and chronic wounds who were repaired with split-thickness skin graft in the Department of Burn and Plastic Surgery of Zhongshan People’s Hospital from September 2021 to September 2022. Ten days after wound healing in the donor area of the thigh (about 4 weeks after surgery), the patients were randomly divided into two groups: the experimental group and the control group. The experimental group was injected with BTX-A solution at the skin donor site by equal interval injection method 1 and 3 months after surgery, 0.1 ml (1 U) was injected at each point, and the interval of each point was 1 cm. The control group was injected with equal amount of normal saline at the skin donor area by equal spacing method 1 and 3 months after operation. The adverse reactions was observed. Follow-up was performed six months after treatment. Evaluation indicators included the doctor’s Vancouver scar scale (VSS) score (including color, vascular distribution, softness and thickness), the visual analogue scale (VAS) score of the patient’s scar pain and itching degree. Both VSS scores and VAS scores were in line with normal distribution, expressed as Mean±SD, and analyzed by independent sample t-test. Results:A total of 60 subjects were recruited, 30 in the experimental group and 30 in the control group. In the experimental group, there were 16 males and 14 females, aged (32.7±5.4) years (18-60 years). In the control group, there were 13 males and 17 females, aged (31.4±4.8) years (18-55 years). There were no significant differences in gender composition and age between the two groups (all P > 0.05). No serious adverse reactions occurred in the experimental group during and after treatment. At the follow-up six months after treatment, the total score of VSS in the experimental group [(3.57±0.60) points vs. (8.52±0.84) points] and the scores of scar color, vascular distribution, flexibility and thickness[(0.88±0.22) points vs. (2.30±0.52) points; (0.73±0.27) points vs. (2.16±0.40) points; (1.29±0.39) points vs. (2.49±0.39) points; (0.66±0.23) points vs. (1.56±0.34) points] were significantly lower than those in the control group, and the differences were statistically significant ( P < 0.01). The VAS score of scar pain and itching degree in the experimental group was also significantly lower than that in the control group [(1.06±0.34) points vs. (2.92±0.63) points], and the difference was statistically significant ( P < 0.01). Conclusion:BTX-A injection is safe and effective in preventing and treating cicatrix in the donor area of split-thickness skin graft, and also has a good effect on relieving the pain and itching symptoms of cicatrix patients.