Objective: To evaluate the inter-observer agreement of the severity of oral epithelial dysplasia in oral leukoplakia, providing a theoretical basis for the development of a more objective grading system. Methods: This study included 60 digital pathological slides of oral leukoplakia from Oral Medicine Department of West China Hospital of Stomatology, Sichuan University, and 239 tissue microarray images of oral leukoplakia from State Key Laboratory of Oral Diseases, Sichuan University, to evaluate the agreement of grading. Besides, 1 000 patches were generated from the 60 digital pathological slides and were divided into 500 small-sized patches (224 pixel×224 pixel) and 500 large-sized patches (1 024 pixel×1 024 pixel), to evaluate the agreement of feature detection. Gradings and feature detections were completed by three pathological experts from the oral pathology departments of two Grade 3, Class A stomatological hospitals in China. Kappa coefficient was used to quantify the inter-observer agreement among pathologists. Results: Minimal agreement was found in the grading of oral epithelial dysplasia among pathologists (Kappa=0.30 in the pathological slide group, Kappa=0.30 in the tissue microarray group). None agreement was found in feature detection within the small-sized patches group (median Kappa=0.14 for architectural features, median Kappa=0.18 for cytological features), and minimal agreement was found in feature detection within the large-sized patches group (median Kappa=0.25 for architectural features, median Kappa=0.25 for cytological features). Conclusions: Generally, the agreement of grading and feature detection of oral epithelial dysplasia in oral leukoplakia is poor. Development of a more objective grading system of oral epithelial dysplasia based on artificial intelligence may be helpful to improve the agreement.
Artificial Intelligence ; China ; Humans ; Leukoplakia, Oral ; Observer Variation ; Precancerous Conditions

Objective:To analyze the liver pathological characteristics of chronic hepatitis B (CHB) patients with normal alanine aminotransferase (ALT) and negative hepatitis B e antigen (HBeAg), and to evaluate the diagnostic value of different serological models for liver fibrosis.Methods:Retrospective analysis was conducted on the patients with HBeAg-negative CHB who had normal ALT and underwent liver biopsy from August 2016 to December 2019 in the Department of Infectious Diseases, Henan Provincial People′s Hospital. The clinical data, serum indicators of hepatitis B virus (HBV) and HBV DNA were collected. The liver fibrosis stages (S) was assessed by pathological examination. The diagnostic efficacies of gamma-glutamyl transpeptidase to platelet ratio (GPR), fibrosis 4 score (FIB-4), S index, aspartate aminotransferase to platelet ratio index (APRI) and gamma-glutamyl transpeptidase to albumin ratio (γ-GT/ALB) for liver pathological fibrosis were analyzed by the receiver operating characteristic curves. Two variable correlation test was used to explore the relationship between the different models and pathological fibrosis of liver tissue. Chi-square test was used for statistical comparison.Results:The age of 448 patients was (37.98±9.82) years, and the male to female ratio was 1.286 ∶1. The proportions of S≥2 in patients with age>30 years, hepatitis B surface antigen (HBsAg)<2 000 IU/mL and HBV DNA≥2 000 IU/mL were higher than those in patients with age ≤30 years, HBsAg ≥2 000 IU/mL and HBV DNA<2 000 IU/mL, respectively, and the differences were all statistically significant ( χ2=7.68, P=0.006; χ2=11.44, P=0.001; χ2=9.12, P=0.003, respectively). There were 250 cases with pathological fibrosis stage S<2, 162 cases with S=2 and 36 cases with S≥3. FIB-4 (correlation coefficient 0.250), APRI (correlation coefficient 0.218), GPR (correlation coefficient 0.186), S index (correlation coefficient 0.184) and γ-GT/ALB (correlation coefficient 0.127) were positively correlated with the severity of liver fibrosis (all P<0.050). S index had the highest sensitivity (64.1%) in the diagnosis of significant liver fibrosis (S≥2), while γ-GT/ALB had the highest specificity (80.8%). In the diagnosis of severe liver fibrosis (S≥3), γ-GT/ALB had the highest sensitivity (77.8%), while APRI had the highest specificity (78.6%). Conclusions:The incidence of liver fibrosis in CHB patients with normal ALT and negative HBeAg is relatively high. The current serological diagnostic models are not suitable for the evaluation of liver fibrosis in these patients, and timely liver puncture is still necessary.

Objective:To analyze the hepatic pathological inflammation and fibrosis condition in order to explore the relationship with related clinical indicators in patients with chronic hepatitis B patients with normal alanine aminotransferase (ALT).Methods:721 cases of chronic hepatitis B with normal ALT who were initially diagnosed in the Department of Infectious Diseases of Henan Provincial People's Hospital from August 2016 to December 2019 were retrospectively collected. Liver biopsy was performed in all patients. General data of patients such as gender, age, liver function indexes, blood routine indexes, HBsAg level, HBeAg status, HBV DNA level, spleen thickness and prothrombin time were collected. Univariate and multivariate analysis methods were used to determine the influencing factors of inflammation and fibrosis degree with liver biopsy. A receiver operating characteristic curve (ROC) was used to evaluate the established multi-factor prediction model. Alpha=0.05 was considered as a standard orientation of test.Results:The average age of 721 cases with chronic hepatitis B was 36.1±9.7 years, and the male to female ratio was 1.28/1, with inflammation and fibrosis grade mainly concentrated in G1S1 (349 cases), G1S2 (132 cases), G2S2 (119 cases), and G2S1 (57 cases). Among them, there were 349 (48.4%) cases of G1S1, and 372 (51.6%) cases of G/S≥2. The main manifestations were mild to moderate inflammation and fibrosis, and only 64 (8.88%) cases had severe G/S≥3. HBsAg level (stratified with 4 log10 IU/ml as the boundary) analyzed in 721 cases were correlated with the relevant clinical indicators stratification and liver pathological inflammation and fibrosis, and the difference was statistically significant (inflammation grade, χ2=6.182, P=0.013; Fibrosis grade, χ2=36.534, P=0.001). Univariate analysis of the relevant clinical indicators that may influence the patient's liver pathological G/S ≥2 showed the patient's age, albumin, γ- glutamyltransferase (GGT), platelet, prothrombin time (PT), spleen thickness and HBsAg level were all statistically significant ( P<0.05), while multivariate analysis showed that age, GGT, PT, and spleen thickness had statistical differences ( P<0.05). The prediction model was established in accordance to multivariate analysis, and the area under the ROC curve was 0.642. Maximization of the sum of sensitivity and specificity as cut-off value of Logit P=0.497, the diagnostic sensitivity, specificity, and Youden's index were 60.6%, 64.5%, and 0.252, respectively. Conclusion:More than half of patients with chronic hepatitis B with normal ALT have significant inflammation and fibrosis and require timely antiviral therapy. Age, GGT, PT and spleen thickness can help comprehensively evaluate the liver inflammation and fibrosis status among patients, but the lack of accurate prediction models suggests that more effective indicators that can help predict the inflammation and fibrosis status of such patients have yet to be discovered. Therefore, liver biopsy should still be actively performed in patients with normal ALT to confirm the diagnosis and timely treatment.

ObjectiveTo investigate the status of abnormal renal function markers and related influencing factors in chronic hepatitis B (CHB) patients receiving oral antiviral drugs for a long time. MethodsA retrospective analysis was performed for the clinical data of 681 CHB patients who attended Henan Provincial People’s Hospital from January to December 2019 and received long-term oral administration of entecavir (ETV)/tenofovir disoproxil fumarate (TDF). All patients received the measurement of blood renal function markers (urea, creatinine, retinol-binding protein ［RBP］, cystatin C ［Cys-C］, and β2-microglobulin ［β2-MG］), urinary renal function markers (α1-microglobulin ［α1-MG］, Cys-C, and N-acetyl-β-D-glucosaminidase ［NAG］), and urine routine parameters. The incidence rate of abnormal renal function markers were analyzed. The McNemar’s test was used for comparison of categorical data between groups, and the multivariate logistic regression analysis was used to investigate independent influencing factors for abnormal renal markers in urine. ResultsThe 681 patients had a mean age of 39.8±11.0 years and received medication for 1.88 (0.80-3.16) years. There were 417 male patients and 264 female patients, and the incidence rate of liver cirrhosis was 27.02% (184/681). Of all 681 patients, 442 received ETV and 239 received TDF. The measurement of blood renal function markers showed that urea, creatinine, retinol-binding protein, Cys-C, and β2-MG had an abnormal rate of 6.9% (47/681), 0.15% (1/681), 0(0/681), 2.21% (15/681), and 5.03% (30/681), respectively, and the abnormal rate of urinary protein was 7.29% (49/672). The measurement of urinary renal function markers showed that α1-MG, NAG, and Cys-C had an abnormal rate of 38.62% (263/681), 37.74% (257/681), and 19.38% (132/681), respectively. The abnormal rate of urine test was higher than that of blood test (P＜0.001). The multivariate logistic regression analysis with urinary α1-MG as the dependent variable showed that sex (odds ratio ［OR］=0.293, 95% confidence interval ［CI］: 0.204-0.419, P＜0.05), age (OR=1298, 95%CI: 1.108-1.521, P＜0.05), and type of nucleoside drug (OR=2.100, 95%CI: 1.431-3.083, P＜0.05) were influencing factors. The multivariate logistic regression analysis with urinary NAG as the dependent variable showed that age (OR=1.177, 95%CI: 1.008-1.375, P=0.040) was an influencing factor. ConclusionCompared with blood renal function markers, urinary renal function markers can identify renal injury earlier in CHB patients, and the elderly patients and the patients receiving TDF are more likely to develop abnormal renal function. However, it is not observed whether the duration of medication and liver cirrhosis can increase the risk of renal injury in CHB patients.

Objective:To evaluate the efficacy and safety of elbasvir/grazoprevir (EBR/GZR) in patients with genotype 1 chronic hepatitis C in the real-world.Methods:This was an open-label, single-center, retrospective real-world study. A total of 103 genotype 1 chronic hepatitis C patients who were treated with EBR/GZR in Henan Provincial People′s Hospital from May 2018 to October 2019 were enrolled.And the clinical baseline characteristics of patients and the effectiveness and safety of antiviral therapy were respectively evaluated.Results:A total of 103 patients were enrolled in the study with an age of (47.6±13.9) years. Fifty-five (53.4%) patients were male and 48(46.6%) were female. One point nine percent (2/103) patients were genotype 1a hepatitis C and 98.1%(101/103) were genotype 1b hepatitis C. Seventeen genotype 1b hepatitis C patients were previously treated with interferon, and three patients co-infected with hepatitis B virus (HBV). Among the 103 cases, 35 had underlying diseases and 26 had combined medication. Ninty-eight cases completed 12-week treatment and 89 cases completed 12-week follow-up after treatment.Overall, 89 cases achieved sustained virological response. The overall incidence of adverse reactions was 20.4%(21/103), and the main adverse reactions were fatigue, insomnia and anxiety. No serious adverse event occurred. The three patients with HBV co-infection had no hepatitis B activation after treatment.Conclusion:EBR/GZR is effective and safe in the patients with genotype 1 chronic hepatitis C in China.

Objective To investigate whether there are differences in lymphocyte subsets between chronic hepatitis B (CHB) patients receiving different antiviral treatment regimens, and to determine related predictive factors for HBsAg decline. Methods A retrospective analysis was performed for 68 treatment-experienced CHB patients who attended the outpatient service in Department of Infectious Diseases, Henan Provincial People's Hospital, from October to December 2019, and according to the antiviral treatment regimen, they were divided into PEG-IFNα treatment group with 10 patients, PEG-IFNα+nucleos(t)ide analogues (NAs) treatment group with 21 patients, and NAs treatment group with 37 patients. Related data were recorded, including demographic features, blood routine, albumin, HBsAg, and measurement of lymphocyte subsets. A one-way analysis of variance was used for comparison of normally distributed continuous data between groups, and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups; the multivariate logistic regression analysis was used to investigate independent influencing factors for HBsAg decline. Results There were significant differences between the three groups in HBsAg decline ( H =8.348, P =0.015), absolute value of lymphocytes ( F =4.643, P =0.013), and T lymphocyte count ( F =7.721, P =0.001). The multivariate logistic regression analysis showed that sex (odds ratio [ OR ]=0.227, 95% confidence interval [ CI ]: 0.059-0.878, P =0.032), age ( OR =0.931, 95% CI : 0.868-0.999, P =0.047), antiviral treatment regimen (PEG-IFN-α treatment group vs NAs treatment group: OR =9.600, 95% CI : 1.982-46.498, P =0.005; PEG-IFN-α+NAs treatment group vs NAs treatment group: OR =4.800, 95% CI : 1.336-17.243, P =0.016), and T lymphocyte count ( OR =0.804, 95% CI : 0.684-0.944, P =0.008) were independent influencing factors for HBsAg decline. Conclusion For CHB patients receiving PEG-IFNα alone or in combination with NAs, monitoring of lymphocyte subsets during the treatment process may help to judge HBsAg decline, and the lower the absolute value of T lymphocytes, the greater the possibility of HBsAg decline.

Objective:To investigate the changes of bone mineral density and its related influencing factors in chronic hepatitis B patients treated with long-term entecavir monotherapy.Methods:211 cases with chronic hepatitis B treated with entecavir monotherapy in the Department of Infectious Diseases of Henan Provincial People’s Hospital from June 2018 to September 2019 were retrospectively collected. Age, gender, body mass index, number of years of medication use, presence or absence of liver cirrhosis and current bone mineral density level (using dual-energy X-ray detection, taking lumbar L1 ~ 4 and left femur as observation region) and other related data were collected. 211 cases general situation was descriptively analyzed by case-control study design. Two independent sample t-tests were used to compare the differences in serum calcium, phosphorus, and renal function levels in patients with different medication durations. Univariate logistic regression was used to screen the influencing factors of bone mineral density level. Significant variables of univariate analysis were included in multivariate logistic regression to obtain the independent influencing factors leading to the decrease of bone mineral density level. The test level was set as α = 0.05.Results:The average age of 211 cases with chronic hepatitis B was (42.36 ± 11.10) years. The average medication time use was (2.52 ± 1.94) years. The body mass index (23.95 ± 3.11), and male-to-female ratio was 2.25/1. The incidence of liver cirrhosis was 35.5%. The incidence of low bone mass in the two observation sites (lumbar spine L1~4 and left femur) was 24.6% and 29.4%, respectively. There were statistically significant differences in serum calcium, phosphorus and renal function levels among patients with different entecavir treatment duration (≥3 years and < 3 years) ( P < 0.05). Univariate analysis result showed that the influencing factors of BMD were age, the number of years of medication use, gender, liver cirrhosis (L1~4 of the lumbar spine region) and age, the number of years of medication, and gender (left femoral region). The variables that entered the two models after the multivariate analysis were age (L1~4 region of lumbar spine: OR = 2.225, left femur OR = 1.660), gender (L1~4 region of lumbar spine: OR = 3.048, left femur OR = 2.496), number of years of medication use (L1~4 region of lumbar spine: OR = 1.387, left femur OR = 1.276). Conclusion:Age, gender, and the number of years of medication use are independent factors that influence the bone mineral density of patients with chronic hepatitis B treated with long-term entecavir. Low bone mass risk at the two observation sites is 2.225 and 1.66 times the normal level for every 10 years of age increase. Compared with men, the risk of low bone mass at the two observation sites is 3.048 and 2.496 times for women, and for every additional year of medication use, the risk of low bone mass at the two observation sites is 1.387 and 1.276 times the normal level. Female patients with older age and prolonged medication use are at high risk of developing bone mineral density reduction.

Objective To investigate the characteristics of blood routine in 129 patients with COVID-19, and analyze the correlation between blood routine parameter changes and clinical classification. Methods A total of 129 COVID-19 patients were recruited and their blood samples were collected at the beginning and the end of treatment.The neutrophil-to-lymphocyte ratio(NLR), platelet-to-lymphocyte ratio(PLR), lymphocyte-to-monocyte ratio(LMR) and C-reactive protein(CRP) were determined and used to calculate Δ NLR, Δ PLR, Δ LMR.The differences in these parameters were compared between the non-severe group (93 cases) and the severe group (36 cases).In addition, the relationship between the changes in blood routine test result and the prognosis of patients was determined. Results The average age of 129 patients was 46.9±17.4 years old, and the ratio of male to female was 1.2 ∶ 1.Thirty-five (35) cases (27.1%) had leucopenia (< 4×10⁹/L) and 59 cases (45.7%) had lymphopenia (< 1.1×10⁹/L).There were statistically significant differences in age, treatment days, blood routine indexes between these two groups.In all the patients, the differences between before and after treatment were statistically significant in the following: leukocyte number, neutrophil cell percentage, lymphocyte number, lymphocyte percentage, monocyte number, monocyte percentage.The differences in RBC, HGB, CRP, and NLR between the two groups before and after treatment were also statistically significant (P < 0.05). Conclusion The older COVID-19 patients have lower number of lymphocytes, higher NLR and PLR, lower LMR and higher CRP.They have a higher risk of progressing to severe disease.After treatment, there is an increase in the number of granulocytes, especially the number of lymphocytes, while a decrease in CRP and NLR.The change of lymphocyte count, NLR and CRP levels can predict the risk of severe COVID-19 and evaluate the therapeutic effect.

Objective:To evaluate the efficacy of artificial liver blood purification system in the treatment of severe and critical patients with corona virus disease 2019(COVID-19), and to observe the dynamic changes of lymphocyte subsets and cytokines after treatment.Methods:A total of six patients with COVID-19 admitted to the ward of public health center of Henan Provincial People′s Hospital from January 31 to February 18, 2020 were enrolled, including three severe cases and three critical cases. The protocals of artificial liver treatment were developed according to the patients′ conditions, and the patients′ epidemiological history, clinical characteristics, and laboratory examination data were retrospectively analyzed. At the same time, dynamic changes of lymphocyte subsets and cytokines were detected before and after artificial liver treatment.Results:By February 24, 2020, two severe patients were discharged after cured, and one case was discharged after improved, with an average stay of 19 days. Two critical patients were still hospitalized and one died. Three severe patients were all treated with hemofiltration, while two critical patients were treated with hemofiltration plus plasma exchange, and one was treated with continuous bedside hemofiltration.Among six patients, the ratio of neutrophils to lymphocytes before treatment were 6.42, 2.63, 15.00, 15.09, 12.04 and 30.41, respectively. After treatment, the ratio of neutrophils to lymphocytes became 4.77, 6.05, 4.86, 5.43, 32.77 and 23.46, respectively.The absolute numbers of lymphocytes in six patients before treatment were low, with a median of 382/μL. After treatment of artificial liver, the absolute numbers of lymphocytes increased, with a median of 476/μL. Cytokines were detected in three critical patients, and the interleukin 6 (IL-6) levels in two cases were 26 042.00 ng/L and 282.03 ng/L, respectively before treatment. After treatment, the levels decreased to 226.85 ng/L and 26.15 ng/L, respectively. IL-6 continued increasing from 30.14 ng/L to 709.25 ng/L in one another critical patient, who eventually died.Conclusions:In severe and critical patients with COVID-19, artificial liver treatment can reduce inflammation and increase the absolute numbers of lymphocytes and the subsets. The IL-6 level may be correlated with disease progression and may be a useful prognostic factor for early identification of severe and critical COVID-19.

Objective:To study whether gene mutation pattern of Gilbert’s syndrome (GS) is combined with viral hepatitis and its relationship with relevant clinical data.Methods:Clinical data of GS patients combined with viral hepatitis who was admitted to the Department of Infectious Diseases of Henan Provincial People's Hospital from August 2013 to December 2018 was retrospectively analyzed. The relationship between gene mutation pattern, general data (age, gender, etc.) and liver biochemical indexes was analyzed. The differences of the above data in patients with or without combined viral hepatitis were analyzed. The measurement data were compared by t-test. The categorical data was compared by the χ2 test. The median and interquartile range of non-normally distributed data was used to indicate the central and discrete tendency. Results:A total of 107 GS eligible cases data were collected. The male to female ratio was 4.94:1 (89:18). The average age of onset was (36.36 ± 12.51) years. Alanine aminotransferase and total bilirubin levels were normal or slightly elevated, while aspartate aminotransferase, alkaline phosphatase, and γ-glutamyltransferase were all within the normal range. There were 49 cases in the combined viral hepatitis group (36 cases with HBV and 13 cases with HCV), and 58 cases in the GS alone group. Total bilirubin level in GS alone group was higher than the combined viral hepatitis group ( z = 0.035, P < 0.05), and there were no statistically significant differences in gender, age, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma glutamyltransferase ( P > 0.05). Uridine diphosphate glucuronide transferase 1A1 (UGT1A1), specifically encoded by GS was detected in all 107 cases. Mutations was mainly occurred in the upstream promoter PBREM-3263 (-3279) (86 cases) and TATA box TA insertion mutation (71 cases), and GGA-AGA Gly71Arg (57 cases) mutation in EXON1 of the coding region. All mutation forms had manifestations of homozygous and heterozygous abnormalities. The combined incidence of main mutation forms in the genetic testing data were sequenced as: A2 + B2 + C2 (17 cases, 25.23%), A1 + B1 (17 cases, 15.89%), A2 (11 cases, 10.28%), C2 (10 Cases, 9.34%), A2 + B2 (7 cases, 6.54%), A1 + B2 (7 cases, 6.54%), C1 (7 cases, 6.54%), and there was no statistically significant difference between different mutation combinations in patients with or without hepatitis ( P > 0.05). The results of total data analysis showed that the total bilirubin level in the single-site mutation group was higher than the multi-site mutation group (Z=2.019, P = 0.043), and other biochemical indicators had no effect ( P > 0.05) and the differences were not statistically significant. Further analysis showed that the total bilirubin level of the single-site mutation subgroup in the GS alone group was higher than the multi-site mutation subgroup ( Z = 1.999, P = 0.046), and the statistical difference was similar to the combined viral hepatitis group ( P > 0.05). Different mutation combinations had no effect on biochemical indexes, and had no relationship with combined viral hepatitis ( P > 0.05). Conclusion:GS is common in patients with combined viral hepatitis, and there is no significant difference between the incidence of gene mutation, mutation forms, biochemical indexes, and non-hepatitis group. The increase in the number of GS mutation sites does not aggravate the deterioration of bilirubin levels due to the decrease in the content and activity of uridine diphosphate glucuronosyltransferase, and the combination of different mutation sites does not affect the changes of various biochemical indexes, and at the same time it is not related to hepatitis.