BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.

AIM: To explore the clinicopathological and immunohistochemistry(IHC)characteristics of meibomian gland carcinoma(MGC).METHODS: Patients who were pathologically diagnosed as MGC from January 1, 2015 to December 31, 2020 in our hospital were enrolled, and their clinicopathological information was retrospectively analyzed. Cancer tissues from all the cases were IHC stained. En Vision two-step method, DAB staining, as well as hematoxylin re-staining were applied in the IHC assay.RESULTS: A total of 50 patients with 21 males and 29 females(1:1.38)were enrolled in the study, ranging from 26 to 80 years old, with a median age of 60 years. The upper eyelid, which was the predilection site, accounting for 66%(33/50). Histopathologically, moderately or poorly differentiated was in the majority(35/50, 70%). The expression rates of IHC parameters of MGC patients were as follows: GATA-3(49/50, 98%), EMA(49/50, 98%), CAM5.2(42/50, 84%), AR(41/50, 82%), MSH2(50/50, 100%), MSH6(50/50, 100%), MLH1(50/50, 100%), PMS2(50/50, 100%), Ki67(positive, 50%-90%). All the patients were followed up for 12 to 72 mo, with 5 cases of recurrence and 0 deaths.CONCLUSION: Pathological diagnosis of MGC should focus on observing cancer cells' cytoplasm to find relevant clues for cortical gland differentiation. Comprehensive analysis of multiple indicators is required when using IHC to assist diagnosis. For most MGC cancer cells, positive expressions of GATA-3, EMA, AR, CAM5.2 and a high Ki67 proliferation index could be always found. In addition, screening for Muir-Torre syndrome related IHC indicators could be also performed in diagnosing MGC simultaneously.

Objective To explore the influencing factors of visual acuity recovery in patients with high myopia after posterior chamber phakic intraocular lens(ICL)implantation.Methods A prospective study was conducted on 210 pa-tients(420 eyes)with high myopia who underwent ICL implantation at the Second Affiliated Hospital of Zhengzhou Univer-sity from May 2021 to March 2023.The patients were divided into a good recovery group[best corrected visual acuity(BC-VA)recovery ≥0.3 D]and a poor recovery group(BCVA recovery＜0.3 D)based on their visual acuity recovery status three months after surgery.The baseline data of patients in the two groups were compared,and the factors affecting visual acuity recovery were analyzed using Logistic regression.The receiver operating characteristic(ROC)curve was used to an-alyze the predictive value of the Logistic regression model for poor visual acuity recovery in patients with high myopia after ICL implantation.Results Three months after surgery,149 patients(298 eyes)were in the good recovery group,and 61 patients(122 eyes)were in the poor recovery group.There were no significant differences in gender,age,years of myopi-a,body mass index,and academic performance between the two groups(all P＞0.05).The proportions of patients with corneal astigmatism＜1.30 D(55.74％),corneal diopter＜45 D(59.02％),Pittsburgh Sleep Quality Index(PSQI)＜7 points(63.93％),and average central radius of curvature[(7.82±0.27)mm]in the poor recovery group were lower than those in the good recovery group[83.89％,81.88％,85.91％,and(7.90±0.24)mm,respectively].The central flat me-ridian curvature(k1)of the anterior corneal surface[(43.27±1.43)D],steep meridian curvature(k2)of the anterior corneal surface[(44.84±1.53)D],and arch height[(628.49±67.28)μm]in the poor recovery group were higher than those in the good recovery group[(42.73±1.42)D,(44.12±1.47)D],and[(417.56±80.14)pm],with significant differences(all P＜0.05).Logistic regression analysis showed that corneal astigmatism,corneal diopter,k1,k2,arch height,and PSQI score were independent influencing factors of poor visual acuity recovery after ICL implantation in pa-tients with high myopia(all P＜0.05).ROC curve analysis showed that the area under the ROC curve for predicting poor visual acuity recovery after ICL implantation in patients with high myopia by Logistic regression model was 0.938(95％CI:0.896-0.966),the sensitivity was 83.61％,and the specificity was 91.95％(P＜0.05).Conclusion The visual acuity recovery after ICL implantation in patients with high myopia is affected by corneal astigmatism,corneal diopter,k1,k2,arch height,and PSQI score.The Logistic regression model based on these factors has high predictive value for visual acui-ty recovery after ICL implantation.

Objective:To assess the diagnostic value of anti-salivary gland protein-1(SP1)antibody combined with anti-parotid secretory protein(PSP)antibody for Sj?gren's syndrome(SS).Methods:A total of 60 patients with primary SS(pSS)who were treated in the outpatient and inpatient department of Department of Rheumatology and Immunology of the Second Hospital of Hebei Medical University from January 2020 to December 2022 were collected.Thirty patients with other autoimmune diseases accompa-nied by dry mouth and/or dry eyes were collected as disease control group.Thirty healthy subjects from the physical examination center were collected for healthy control group,serum samples were obtained from all of them.Their general features and clinical information including clinical manifestations,labora-tory examinations and other examinations were recorded.The 2016 American College of Rheumatology(ACR)/European League against Rheumatism(EULAR)classification criteria were adopted as the diag-nostic standard of pSS.Immunoglobulin G(IgG)subtype of anti-SP1 antibody and anti-PSP antibody were detected by chemiluminescence immunoassay.The receiver operating characteristic(ROC)curve was used to evaluate the accuracy of anti-SP1 antibody and anti-PSP antibody in diagnosing pSS.The cli-nical characteristics of anti-SP1 antibody and anti-PSP antibody positive patients and negative patients in pSS group were further compared.Independent samples t test,Mann-Whitney U test,variance analysis,Kruskal-Wallis test,Chi-square test or Fisher's exact test and Spearman correlation analysis were used for statistical analysis.Results:There was no significant difference in age(F=1.406,P=0.495)and gender(x2=2.105,P=0.349)among pSS group,disease control group and healthy control group.The expression levels of anti-SP1 antibody(H=16.73,P＜0.001)and anti-PSP antibody(H=26.09,P＜0.001)were statistically different among the three groups.An intergroup comparison of anti-SP1 antibody expression levels showed that there was a statistically significant difference between pSS and healthy con-trol group(P＜0.001),but no statistically significant difference between the other groups.Comparison of anti-PSP antibody expression levels between the groups showed that there were statistically significant differences between pSS and healthy control group(P＜0.001),and between disease control group and healthy control group(P=0.009),while no statistically significant differences between the other groups.The positive rate of anti-SP1 antibody in pSS group was significantly higher than that in disease control group and healthy control group(58.33％vs.40.00％vs.13.33％,P＜0.001).The positive rate of anti-PSP antibody in pSS group was significantly higher than that in disease control group and healthy control group(75.00％vs.56.17％vs.16.67％,P＜0.001).The area under the curve for anti-SP1 antibody was 0.688(P＜0.001).The sensitivity and specificity of anti-SP1 antibody were 58.33％(35/60)and 70.00％(42/60)respectively,the positive predictive value was 66.04％(35/53)and the negative predictive value was 54.55％(42/77)of anti-SP1 antibody.The area under the curve of anti-PSP antibody was 0.720(P＜0.001),with a sensitivity was 75.00％(45/60),and spe-cificity was 63.33％(38/60).The positive predictive value and negative predictive value of anti-PSP an-tibody were 67.16％(45/67)and 71.70％(38/53)respectively.All the 13 pSS patients were negative for anti-Sjogren's syndrome A(SSA,including SSA52 and SSA60)antibody and anti-Sjogren's syn-drome B(SSB)antibody.Among them,11 patients were positive for both anti-SP1 antibody and anti-PSP antibody,1 patient was positive for anti-SP1 antibody and 1 patient was positive for anti-PSP anti-body.The clinical features of anti-SP1 antibody and anti-PSP antibody positive and negative groups were compared in pSS patients.The duration of disease in anti-SP1 antibody positive group was shorter(Z=-2.277,P=0.023)when compared with the negative patients.The patients with positive anti-PSP an-tibody were younger than those in the negative group(t=2.598,P＜0.05),the positive rate of rheuma-toid factor(P=0.002)and the serum level of IgG(t=3.806,P=0.003)in anti-PSP antibody positive group were higher than in the negative group.Analysis of the correlation between anti-SP1 antibody and anti-PSP antibody in the pSS patients showed that there was significant correlation between them(r=0.801,P＜0.001).Conclusion:Both anti-SP1 antibody and anti-PSP antibody are valuable in the diag-nosis of SS,and anti-SP1 antibody is helpful for the early diagnosis of pSS.The combined detection of anti-SP1 antibody and anti-PSP antibody is helpful for the early diagnosis of pSS patients with negative anti-SSA antibody and anti-SSB antibody.

Traditional Chinese medicine (TCM) has played an important role in the prevention and treatment of Coronavirus disease 2019 (COVID-19) epidemic in China. The integration of Chinese and Western medicine is an important feature of Chinese COVID-19 prevention and treatment. According to a series of evidence-based studies, TCM can reduce the infection rate of severe acute respiratory syndrome coronavirus 2 in high-risk groups. For patients with mild and moderate forms of COVID-19, TCM can relieve the related signs and symptoms, shorten the period of nucleic-acid negative conversion, and reduce conversion rate to the severe form of the disease. For COVID-19 patients with severe and critical illnesses, TCM can improve inflammatory indicators and blood oxygen saturation, shorten the hospital stay, and reduce the mortality rate. During recovery, TCM can improve patients' symptoms, promote organ function recovery, boost the quality of patients' life, and reduce the nucleic-acid repositive conversion rate. A series of mechanism research studies revealed that capability of TCM to treat COVID-19 through antiviral and anti-inflammatory effects, immune regulation, and protection of organ function via a multicomponent, multitarget, and multipathway approach.
Humans ; COVID-19 ; Medicine, Chinese Traditional ; Drugs, Chinese Herbal/therapeutic use* ; SARS-CoV-2 ; Epidemics

Objective:To investigate the protective effect and possible mechanism of Tangshenbao on renal damage in diabetic nephropathy (DN) rats.Methods:Totally 36 SPF male SD rats were randomly divided into normal group ( n=6) and model group ( n=30). The DN rat model was prepared by single high-dose intraperitoneal injection of STZ. According to the random number table method, the rats were divided into model group, irbesartan group and Tangshenbao low-, medium- and high-dosage groups, with 6 rats in each group. Drug intervention lasted for 8 weeks. The general condition and body weight of rats in each group were recorded. The blood glucose, kidney index, 24 h urine protein (24 h UTP), SCr and BUN levels were detected. The pathological morphology of renal tissue was observed by PAS staining and transmission electron microscopy. The mRNA and protein expressions of Ets-1, TGF-β1, Smad2 and Smad3 in renal tissue were detected by real-time fluorescence quantitative PCR and Western blot. Results:Compared with model group, the body weight of Tangshenbao low, medium and high dose groups and irbesartan group significantly increased ( P<0.01). The kidney index decreased ( P<0.05 or P<0.01). The contents of 24 hUTP, BUN and SCr significantly decreased ( P<0.05 or P<0.01). Glomerular volume was significantly reduced ( P<0.05 or P<0.01), the mRNA expressions of Ets-1 (1.59 ± 0.06, 1.47 ± 0.04, 1.31 ± 0.03, 1.39 ± 0.03 vs. 1.64 ± 0.04), TGF-β1 (1.65 ± 0.05, 1.59 ± 0.03, 1.38 ± 0.05, 1.49 ± 0.04 vs. 1.77 ± 0.08), Smad2 (1.48 ± 0.05,1.39 ± 0.05, 1.22 ± 0.03, 1.31 ± 0.04 vs. 1.54 ± 0.05), Smad3 (1.57 ± 0.04, 1.48 ± 0.03, 1.28 ± 0.03, 1.39 ± 0.02 vs. 1.64 ± 0.05) in renal tissue of rats significantly decreased ( P<0.05 or P<0.01), the protein expressions of Ets-1 (1.33 ± 0.32, 1.16 ± 0.38, 0.77 ± 0.06, 0.84 ± 0.06 vs. 1.97 ± 0.43), TGF-β1 ( 1.35 ± 0.14, 1.24 ± 0.22, 0.94 ± 0.13, 1.07 ± 0.06 vs. 1.63 ± 0.20), Smad2 (1.24 ± 0.26, 1.14 ± 0.31, 0.77 ± 0.28, 0.85 ± 0.19 vs. 1.72 ± 0.34) and Smad3 (1.29 ± 0.14, 1.19 ± 0.21, 0.85 ± 0.39, 0.90 ± 0.37 vs. 1.76 ± 0.21) decreased ( P<0.05 or P<0.01). Conclusion:Tangshenbao can improve renal damage in DN rats, and its mechanism may be related to the inhibition of Ets-1 expression and TGF-β1/Smads signaling pathway.

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)-induced cytokine storms constitute the primary cause of coronavirus disease 19(COVID-19)progression,severity,criticality,and death.Gluco-corticoid and anti-cytokine therapies are frequently administered to treat COVID-19,but have limited clinical efficacy in severe and critical cases.Nevertheless,the weaknesses of these treatment modalities have prompted the development of anti-inflammatory therapy against this infection.We found that the broad-spectrum anti-inflammatory agent inosine downregulated proinflammatory interleukin(IL)-6,upregulated anti-inflammatory IL-10,and ameliorated acute inflammatory lung injury caused by mul-tiple infectious agents.Inosine significantly improved survival in mice infected with SARS-CoV-2.It indirectly impeded TANK-binding kinase 1(TBK1)phosphorylation by binding stimulator of interferon genes(STING)and glycogen synthase kinase-3β(GSK3β),inhibited the activation and nuclear trans-location of the downstream transcription factors interferon regulatory factor(IRF3)and nuclear factor kappa B(NF-κB),and downregulated IL-6 in the sera and lung tissues of mice infected with lipopoly-saccharide(LPS),H1N1,or SARS-CoV-2.Thus,inosine administration is feasible for clinical anti-inflammatory therapy against severe and critical COVID-19.Moreover,targeting TBK1 is a promising strategy for inhibiting cytokine storms and mitigating acute inflammatory lung injury induced by SARS-CoV-2 and other infectious agents.

Abstract: Objective　To construct HepG2, Huh7 cell lines stably express hepatitis B virus X (HBx) mutant (C1653T, T1753C), and explore their effect on the biological behavior of hepatocellular carcinoma cells. Methods　The lentivirus plasmid of pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were obtained by PCR site mutagenesis according to wild type ayr HBx. Double enzyme digestion and Sanger sequencing were performed for accuracy of plasmid. Blank HepG2 and Huh7 cells were used as the control group, HepG2, Huh7 cells were infected by pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, and pLVX-HBxT1753C-IRES-tdTomato lentivirus solution, then monoclonal cell was selected by 0.6 μg/mL puromycin. Immunostaining and Western Blot were performed for the verification of stable strains. CCK8 assay was performed for the proliferation capacity of stable strains. Western Blot was performed for expression of EMT-related signal molecules in cells. The independent samples t-test was used for comparison between two groups. Results Double enzyme digestion and Sanger sequencing showed that that the size of the cut fragments of recombinant lentiviral plasmids was correct, and the point mutation location and base substitution were correct, suggesting that the plasmid of pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were constructed successfully. Immunostaining and Western blot showed that HBX were expressed in stable strains, while there was no HBX expression in the blank control group, indicating that the HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed. CCK8 assay showed that the proliferation capacity of HBx and mutant were enhanced compared to the control group (P<0.01), HBx C1653T displayed further additive the effect compared to HBx (P<0.05). Moreover, HBxC1653T mutation also significantly upregulated N-cadherin expression and downregulated E-cadherin expression, thus promoting the occurrence of EMT. Conclusions　HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed, HBxC1653T mutation significantly enhanced the proliferation of HCC cells and epithelial to mesenchymal transition occurrence.

Thyroid eye disease (TED) is an autoimmune disease closely related to Graves' disease and also the most common orbitopathy, which remains a challenge to diagnose and manage.In addition to the updated clinical practice guideline released by European Group on Graves' Orbitopathy in 2021 and the Chinese guideline released by Chinese Medical Association in 2022, a consensus statement was made by the American Thyroid Association and the European Thyroid Association in 2022.This consensus statement reviewed the latest research progress and clinical experience summarized from the last few decades as an aid to doctors involved in the management of TED.Further elucidation of key points from the consensus statement will be provided in this paper.

Objective:To investigate the effectiveness of early levothyroxine intervention in pregnancy complicated with hypothyroidism.Methods:A retrospective analysis was performed on 132 pregnant patients with hypothyroidism admitted to the Department of Endocrinology, Anyang People’s Hospital from Nov. 2018 to Sep. 2021. Among them, 68 cases with levothyroxine sodium early intervention were included in the intervention group, and 64 cases without intervention were included in the non-intervention group. The differences in vascular endothelial function indexes, thyroid function indexes, and blood lipid indexes before and after treatment in the intervention group were compared with those in the non-intervention group. The incidence of obstetric complications such as preeclampsia, fetal growth restriction, and adverse pregnancy outcomes of abortion and neonatal asphyxia were compared between the two groups. SPSS 21.0 software was used to process data, measurement data were subjected to t test, and enumeration data were subjected to χ 2 test. Results:After treatment, the endothelium-dependent brachial artery blood flow-mediated vasodilation index (FMD) index, nitric oxide (NO) , and endothelin-1 (ET-1) levels in the intervention group were significantly better than those in the non-intervention group [ (10.37%) ±1.54%) vs (7.25% ± 1.09%) , (60.85 ± 7.03) umol/L vs (39.11 ± 4.31) umol/L, (112.96 ± 13.58) umol/L vs (238.85 ± 26.05) umol/L]. After treatment, the serum thyroid stimulating hormone (TSH) in the intervention group was significantly lower than that in the non-intervention group [ (2.25±0.26) mU/L vs (8.79±1.60) mU/L] ( P<0.001) . After treatment, the levels of total cholesterol (TC) , triacylglycerol (TG) and low-density lipoprotein cholesterol (LDL-C) in the intervention group were significantly lower than those in the non-intervention group [ (3.52±0.91) mmol/L vs (6.51±1.31) mmol/L L, (1.30±0.31) mmol/L vs (1.44±0.36) mmol/L, (2.29±0.31) mmol/L vs (3.32±0.44) mmol/L] ( P<0.001, P=0.036, P<0.001) . The incidence of obstetric complications such as preeclampsia, fetal growth restriction, premature rupture of membranes, and abnormal amniotic fluid volume in the intervention group were significantly lower than those in the non-intervention group (5.88% vs 17.19%, 1.47% vs 9.38%, 10.29% vs. 23.44%, 2.94% vs 12.50%) ( P=0.041, 0.043, 0.043, 0.038) ; the incidence of miscarriage, premature birth, neonatal asphyxia, cesarean section and other adverse pregnancy outcomes in the intervention group were significantly lower than those in the non-intervention group (1.47% vs 9.38%, 4.69% vs 15.36%, 2.94% vs 10.94%, 57.35% vs 75.00%) ( P=0.043, 0.031, 0.038, 0.033) . Conclusion:Early intervention with levothyroxine in pregnancy complicated with hypothyroidism is beneficial to improve thyroid function, reduce blood lipid level, protect vascular endothelial function, reduce related obstetric complications, and reduce the incidence of adverse pregnancy outcomes for mothers and infants.