AIM: To investigate the correlation and diagnostic performance between baseline ocular parameters and the occurrence of retinopathy in patients with type 2 diabetes mellitus(T2DM)without retinopathy.METHODS: From May 2020 to May 2021, our hospital admitted a total of 123 patients with type 2 diabetes mellitus(T2DM)who had no retinopathy at baseline. Clinical data and ocular parameters at baseline were prospectively collected, followed by at least a 3-year follow-up period. The number of cases of diabetic retinopathy occurred during the follow-up period was statistically analyzed, and a multivariable Logistic regression analysis was conducted to identify factors associated with the development of diabetic retinopathy. ROC curve analysis was performed on the factors identified in the multivariable Logistic regression analysis.RESULTS: During the follow-up period, 49 patients(39.8%)developed diabetic retinopathy. Patients who developed retinopathy were included in the retinopathy group, while the remaining patients were included in the non-retinopathy group(n=74). The fasting blood glucose, glycated hemoglobin(HbA1c), anterior chamber depth, and central corneal thickness were higher in the retinopathy group than in the non-retinopathy group(all P<0.01), while the axial length and pupil diameter were lower in the retinopathy group than in the non-retinopathy group(all P<0.01). Low fasting blood glucose, HbA1c, and central corneal thickness are protective factors against diabetic retinopathy in T2DM patients [OR(95% CI): 0.356(0.130-0.978), 0.213(0.057-0.797), 0.971(0.950-0.992), all P<0.05], while low pupil diameter is a risk factor for diabetic retinopathy in T2DM patients [OR(95% CI): 389.483(20.298-7473.596), P<0.05]. ROC curve analysis showed that the AUC was highest when combining fasting blood glucose, HbA1c, central corneal thickness, and pupil diameter, at 0.962(95% CI: 0.934-0.991). The AUC values for the central corneal thickness and pupil diameter were 0.725(95% CI: 0.596-0.814)and 0.711(95% CI: 0.602-0.814), respectively, which were not significantly different from the AUC values of fasting blood glucose 0.709(95% CI: 0.615-0.803)and HbA1c 0.708(95% CI: 0.613-0.804), while the combined diagnosis of central corneal thickness and pupil diameter can increase the AUC to 0.896(95% CI: 0.835-0.976).CONCLUSION: Baseline ocular parameters, including central corneal thickness and pupil diameter, in patients with T2DM can be used to predict the risk of developing diabetic retinopathy.

The differences in chemical quality characteristics between Xinjiang Rehmannia glutinosa and R. glutinosa were analyzed to provide a theoretical basis for the introduction and quality control of R. glutinosa. In this study, the high performance liquid chromatography(HPLC) fingerprints of 6 batches of Xinjiang R. glutinosa and 10 batches of R. glutinosa samples were established. The content of iridoid glycosides, phenylethanoid glycosides, monosaccharides, oligosaccharides, and polysaccharides in Xinjiang R. glutinosa and R. glutinosa was determined by high performance liquid chromatography-diode array detection(HPLC-DAD), high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD), and ultraviolet-visible spectroscopy(UV-Vis). The determination results were analyzed with by chemical pattern recognition and entropy weight TOPSIS method. The results showed that there were 19 common peaks in the HPLC fingerprints of the 16 batches of R. glutinosa, and catalpol, aucubin, rehmannioside D, rehmannioside A, hydroxytyrosol, leonuride, salidroside, cistanoside A, and verbascoside were identified. Hierarchical cluster analysis(HCA) and principal component analysis(PCA) showed that Qinyang R. glutinosa, Mengzhou R. glutinosa, and Xinjiang R. glutinosa were grouped into three different categories, and eight common components causing the chemical quality difference between Xinjiang R. glutinosa and R. glutinosa in Mengzhou and Qinyang of Henan province were screened out by orthogonal partial least squares discriminant analysis(OPLS-DA). The results of content determination showed that there were glucose, sucrose, raffinose, stachyose, polysaccharides, and nine glycosides in Xinjiang R. glutinosa and R. glutinosa samples, and the content of catalpol, rehmannioside A, leonuride, cistanoside A, verbascoside, sucrose, and glucose was significantly different between Xinjiang R. glutinosa and R. glutinosa. The analysis with entropy weight TOPSIS method showed that the comprehensive quality of R. glutinosa in Mengzhou and Qinyang of Henan province was better than that of Xinjiang R. glutinosa. In conclusion, the types of main chemical components of R. glutinosa and Xinjiang R. glutinosa were the same, but their content was different. The chemical quality of R. glutinosa was better than Xinjiang R. glutinosa, and other components in R. glutinosa from two producing areas and their effects need further study.
Rehmannia/classification* ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Quality Control

OBJECTIVE: To screen novel diagnostic marker or therapeutic target for multiple myeloma (MM). METHODS: Sel1L, SPAG4, KCNN3 and PARM1 were identified by bioinformatics method based on GEO database as high expression genes in MM. Their RNA and protein expression levels in bone marrow mononuclear cells from myeloma cell lines U266, NCI-H929, MM.1s, RPMI8226 and leukemia cell line THP1, as well as 31 MM patients were evaluated by RT-PCR and Western blot, respectively. Meanwhile, 5 samples of bone marrow from healthy donors for allogeneic hematopoietic stem cell transplantation were employed as controls. RESULTS: Compared with leukemia cell line THP1, the expression levels of KCNN3, PARM1 and Sel1L mRNA were significantly increased in myeloma cell lines U266, NCI-H929 and MM.1s, while PARM1 was further increased in myeloma cell lines 8226. Western blot showed that the 4 genes were all expressed in the 4 myeloma cell lines. Compared with healthy controls, the expression levels of Sel1L, SPAG4, KCNN3 and PARM1 mRNA were significantly higher in MM patients (all P < 0.05). Western blot showed that the 4 genes were all expressed in MM patients, and the protein expression level of Sel1L and KCNN3 were significantly different compared with healthy donors (all P < 0.01). CONCLUSION Sel1L, SPAG4, KCNN3 and PARM1 may be potential diagnostic markers and therapeutic targets for MM.
Humans ; Multiple Myeloma/genetics* ; Cell Line, Tumor ; Proteins/metabolism* ; Computational Biology ; RNA, Messenger/genetics*

OBJECTIVE: To investigate the therapeutic potential of pseudolaric acid B (PAB) on non-alcoholic fatty liver disease (NAFLD) and its underlying molecular mechanism in vitro and in vivo. METHODS: Eight-week-old male C57BL/6J mice (n=32) were fed either a normal chow diet (NCD) or a high-fat diet (HFD) for 8 weeks. The HFD mice were divided into 3 groups according to a simple random method, including HFD, PAB low-dose [10 mg/(kg·d), PAB-L], and PAB high-dose [20 mg/(kg·d), PAB-H] groups. After 8 weeks of treatment, glucose metabolism and insulin resistance were assessed by oral glucose tolerance test (OGTT) and insulin tolerance test (ITT). Biochemical assays were used to measure the serum and cellular levels of total cholesterol (TC), triglycerides (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). White adipose tissue (WAT), brown adipose tissue (BAT) and liver tissue were subjected to hematoxylin and eosin (H&E) staining or Oil Red O staining to observe the alterations in adipose tissue and liver injury. PharmMapper and DisGeNet were used to predict the NAFLD-related PAB targets. Peroxisome proliferator-activated receptor alpha (PPARα) pathway involvement was suggested by Kyoto Encyclopedia of Genes and Genomes (KEGG) and search tool Retrieval of Interacting Genes (STRING) analyses. Luciferase reporter assay, cellular thermal shift assay (CETSA), and drug affinity responsive target stability assay (DARTS) were conducted to confirm direct binding of PAB with PPARα. Molecular dynamics simulations were applied to further validate target engagement. RT-qPCR and Western blot were performed to assess the downstream genes and proteins expression, and validated by PPARα inhibitor MK886. RESULTS: PAB significantly reduced serum TC, TG, LDL-C, AST, and ALT levels, and increased HDL-C level in HFD mice (P<0.01). Target prediction analysis indicated a significant correlation between PAB and PPARα pathway. PAB direct target binding with PPARα was confirmed through luciferase reporter assay, CETSA, and DARTS (P<0.05 or P<0.01). The target engagement between PAB and PPARα protein was further confirmed by molecular dynamics simulations and the top 3 amino acid residues, LEU321, MET355, and PHE273 showed the most significant changes in mutational energy. Subsequently, PAB upregulated the genes expressions involved in lipid metabolism and mitochondrial biogenesis downstream of PPARα (P<0.05 or P<0.01). Significantly, the PPARα inhibitor MK886 effectively reversed the lipid-lowering and PPARα activation properties of PAB (P<0.05 or P<0.01). CONCLUSION PAB mitigates lipid accumulation, ameliorates liver damage, and improves mitochondrial biogenesis by binding with PPARα, thus presenting a potential candidate for pharmaceutical development in the treatment of NAFLD.
Animals ; PPAR alpha/metabolism* ; Non-alcoholic Fatty Liver Disease/pathology* ; Male ; Mice, Inbred C57BL ; Lipid Metabolism/drug effects* ; Diterpenes/therapeutic use* ; Organelle Biogenesis ; Diet, High-Fat ; Humans ; Mice ; Liver/metabolism* ; Insulin Resistance ; Mitochondria/metabolism* ; Molecular Docking Simulation

OBJECTIVE: Hepatocellular carcinoma (HCC) is sensitive to ferroptosis, a new form of programmed cell death that occurs in most tumor types. However, the mechanism through which ferroptosis modulates HCC remains unclear. This study aimed to investigate the oncogenic role and prognostic value of FANCD2 and provide novel insights into the prognostic assessment and prediction of immunotherapy. METHODS: Using clinicopathological parameters and bioinformatic techniques, we comprehensively examined the expression of FANCD2 macroscopically and microcosmically. We conducted univariate and multivariate Cox regression analyses to identify the prognostic value of FANCD2 in HCC and elucidated the detailed molecular mechanisms underlying the involvement of FANCD2 in oncogenesis by promoting iron-related death. RESULTS: FANCD2 was significantly upregulated in digestive system cancers with abundant immune infiltration. As an independent risk factor for HCC, a high FANCD2 expression level was associated with poor clinical outcomes and response to immune checkpoint blockade. Gene set enrichment analysis revealed that FANCD2 was mainly involved in the cell cycle and CYP450 metabolism. CONCLUSION To the best of our knowledge, this is the first study to comprehensively elucidate the oncogenic role of FANCD2. FANCD2 has a tumor-promoting aspect in the digestive system and acts as an independent risk factor in HCC; hence, it has recognized value for predicting tumor aggressiveness and prognosis and may be a potential biomarker for poor responsiveness to immunotherapy.
Humans ; Carcinoma, Hepatocellular/diagnosis* ; Liver Neoplasms/diagnosis* ; Immunotherapy ; Fanconi Anemia Complementation Group D2 Protein/metabolism* ; Prognosis ; Male ; Female ; Middle Aged ; Biomarkers, Tumor/metabolism*

OBJECTIVE: To investigate chronic hepatitis C virus (HCV) infection's effect on gestational liver function, pregnancy and delivery complications, and neonatal development. METHODS: A total of 157 HCV antibody-positive (anti-HCV[+]) and HCV RNA(+) patients (Group C) and 121 anti-HCV(+) and HCV RNA(-) patients (Group B) were included as study participants, while 142 anti-HCV(-) and HCV RNA(-) patients (Group A) were the control group. Data on biochemical indices during pregnancy, pregnancy complications, delivery-related information, and neonatal complications were also collected. RESULTS: Elevated alanine aminotransferase (ALT) rates in Group C during early, middle, and late pregnancy were 59.87%, 43.95%, and 42.04%, respectively-significantly higher than Groups B (26.45%, 15.70%, 10.74%) and A (23.94%, 19.01%, 6.34%) ( P < 0.05). Median ALT levels in Group C were significantly higher than in Groups A and B at all pregnancy stages ( P < 0.05). No significant differences were found in neonatal malformation rates across groups ( P > 0.05). However, neonatal jaundice incidence was significantly greater in Group C (75.16%) compared to Groups A (42.25%) and B (57.02%) ( χ ² = 33.552, P < 0.001). HCV RNA positivity during pregnancy was an independent risk factor for neonatal jaundice ( OR = 2.111, 95% CI 1.242-3.588, P = 0.006). CONCLUSIONS Chronic HCV infection can affect the liver function of pregnant women, but does not increase the pregnancy or delivery complication risks. HCV RNA(+) is an independent risk factor for neonatal jaundice.
Humans ; Female ; Pregnancy ; Adult ; Pregnancy Complications, Infectious/epidemiology* ; Retrospective Studies ; Pregnancy Outcome ; Infant, Newborn ; Viremia/virology* ; Hepatitis C ; Hepacivirus/physiology* ; Hepatitis C, Chronic/virology* ; Young Adult ; Alanine Transaminase/blood*

Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).

Objective:To study the expression levels of long non-coding RNA(lncRNA)H19 and insulin-like growth factor 2(IGF2)genes in breast cancer tissue,and to analyze their imprinting status.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of H19 and IGF2 mRNA in breast cancer tissue and adjacent tissue,and the differences in the expressions of H19 and IGF2 mRNA in breast cancer tissue and adjacent tissue were analyzed;single nucleotide polymorphism(SNP)was used to distinguish the allele expression status(homozygous or heterozygous).For heterozygous IGF2(Apa Ⅰ site)or H19(Alu Ⅰ site)in genomic DNA,imprinting analysis was used to detect the imprinting status of H19 and IGF2 in breast cancer tissue,that were maintenance of imprinting(MOI)or loss of imprinting(LOI);the relationship between the expressions of H19 and IGF2 and molecular subtypes in breast cancer tissue were also analyzed.Results:The RT-qPCR results showed that the expression levels of H19 and IGF2 mRNA in breast cancer tissue were higher than those in adjacent tissue(P＜0.01).There was a positive correlation between the expression levels of H19 mRNA and IGF2 mRNA(r=0.567,P＜0.01).Compared with adjacent tissue,the expression levels of H19 mRNA in cancer tissue of the breast cancer patients with various molecular subtypes were increased(P＜0.05 or P＜0.01).LOI was observed in both H19 and IGF2 in breast cancer tissue,and the incidence of IGF2 LOI was 36.7％,which was higher than that of H19 LOI(4.3％).The RT-qPCR results showed that the expression level of IGF2 mRNA in breast cancer tissue in IGF2 LOI group was significantly higher than that in IGF2 MOI group(P＜0.01).Conclusion:The expression levels of H19 and IGF2 mRNA in breast cancer tissue are significantly higher than those in adjacent tissue.The incidence of IGF2 LOI is higher than that of H19 LOI,and IGF2 LOI may be one of the key factors in the pathogenesis of breast cancer.

Objective:To explore the effects and mechanism of Bushen Tongluo Prescription in mechanical injury of rat endometrium.Method:A total of 60 female SD rats were divided into the blank group, model group, estradiol valerate group, and Bushen Tongluo Prescription group according to the random number table method, with 15 rats in each group. Except for the blank group, the other three groups were used to establish a rat model of endometrial injury using mechanical injury method. After modeling, Bushen Tongluo Prescription group was orally administered with Bushen Tongluo Prescription decoction at a dosage of 2.1 g/kg, the estradiol group was orally administered with estradiol valerate at a dosage of 0.4 mg/kg, and the blank group and model group were orally administered with 0.5% CMC at an equal dosage, once a day, for a total of 8 days. Samples were taken on the 1st, 4th, and 8th day after gastric lavage. The thickness of the endometrium and glands were observed using HE staining, the degree of uterine tissue fibrosis was observed using Masson staining, and VEGF and TGF-β protein and mRNA expressions in uterine tissue were detected using Western blot and fluorescence quantitative PCR.Results:Compared with the Bushen Tongluo Prescription on the first day, the thickness of the endometrium and the number of glands increased on the eighth day ( P<0.05); compared with the model group, the number of glands in the Bushen Tongluo Prescription group increased on the 4th day of administration ( P<0.05), while the fibrotic area of the endometrium decreased on the 8th day of administration ( P<0.05); compared with the model group, on the 8th day of administration, the expression of VEGF protein and TGF-β1 in the Bushen Tongluo Prescription group increased ( P<0.05), and protein expression decreased ( P<0.05); compared with the model group, the group treated with Bushen Tongluo Prescription had TGFβ1 mRNA level on the first day increased ( P<0.05), while the level of TGF-β1mRNA decreased on the 8th day ( P<0.05). Conclusion:Bushen Tongluo Prescription can increase endometrial thickness and receptivity, effectively improve damaged endometrium, anti fibrotic and prevent endometrial adhesion by up-regulating VEGF protein expression and down-regulating TGF-β1 protein and mRNA expression.