[Clinical features and prognosis of core binding factor acute myeloid leukemia children in South China: a multicenter study].

Objective: To analyze the clinical features, efficacy and prognosis factors of core binding factor (CBF) acute myeloid leukemia (AML) children in South China. Methods: This was a retrospective cohort study. Clinical data of 584 AML patients from 9 hospitals between January 2015 to December 2020 was collected. According to fusion gene results, all patients were divided into two groups: CBF-AML group (189 cases) and non-CBF-AML group (395 cases). CBF-AML group were divided into AML1-ETO subgroup (154 cases) and CBFß-MYH11 subgroup (35 cases). Patients in CBF-AML group chosen different induction scheme were divided into group A (fludarabine, cytarabine, granulocyte colony stimulating factor and idarubicin (FLAG-IDA) scheme, 134 cases) and group B (daunorubicin, cytarabine and etoposide (DAE) scheme, 55 cases). Age, gender, response rate, recurrence rate, mortality, molecular genetic characteristics and other clinical data were compared between groups. Kaplan-Meier method was used for survival analysis and survival curve was drawn. Cox regression model was used to analyze prognostic factors. Results: A total of 584 AML children were diagnosed, including 346 males and 238 females. And a total of 189 children with CBF-AML were included, including 117 males and 72 females. The age of diagnosis was 7.3 (4.5,10.0)years, and the white blood cell count at initial diagnosis was 21.4 (9.7, 47.7)×109/L.The complete remission rate of the first course (CR1) of induction therapy, relapse rate, and mortality of children with CBF-AML were significantly different from those in the non-CBF-AML group (91.0% (172/189) vs. 78.0% (308/395); 10.1% (19/189) vs. 18.7% (74/395); 13.2% (25/189) vs. 25.6% (101/395), all P<0.05). In children with CBF-AML, the CBFß-MYH11 subgroup had higher initial white blood cells and lower proportion of extramedullary invasion than the AML1-ETO subgroup, with statistical significance (65.7% (23/35) vs. 14.9% (23/154), 2.9% (1/35) vs. 16.9% (26/154), both P<0.05). AML1-ETO subgroup had more additional chromosome abnormalities (75/154), especially sex chromosome loss (53/154). Compared with group B, group A had more additional chromosome abnormalities and a higher proportion of tumor reduction regimen, with statistical significance (50.0% (67/134) vs. 29.1% (16/55), 34.3% (46/134) vs. 18.2% (10/55), both P<0.05). Significant differences were found in 5-years event free survival (EFS) rate and 5-year overall survival (OS) rate between CBF-AML group and non-CBF-AML group ((77.0±6.4)%vs. (61.9±6.7)%,(83.7±9.0)%vs. (67.3±7.2)%, both P<0.05).EFS and OS rates of AML1-ETO subgroup and CBFß-MYH11 subgroup in children with CBF-AML were not significantly different (both P>0.05). Multivariate analysis showed in the AML1-ETO subgroup, CR1 rate and high white blood cell count (≥50×109/L) were independent risk factors for EFS (HR=0.24, 95%CI 0.07-0.85,HR=1.01, 95%CI 1.00-1.02, both P<0.05) and OS (HR=0.24, 95%CI 0.06-0.87; HR=1.01, 95%CI 1.00-1.02; both P<0.05). Conclusions: In CBF-AML, AML1-ETO is more common which has a higher extramedullary involvement and additional chromosome abnormalities, especially sex chromosome loss. The prognosis of AML1-ETO was similar to that of CBFß-MYH11. The selection of induction regimen group FLAG-IDA for high white blood cell count and additional chromosome abnormality can improve the prognosis.

[Association of greenness, nitrogen dioxide with the prevalence of hypertension among the elderly over 65 years old in China].

Objective: To investigate the association of mixed exposure to greenness and nitrogen dioxide(NO2) and hypertension among the older adults aged 65 years and over in China. Methods: The study subjects were from the Chinese Longitudinal Healthy Longevity Survey from 2017 to 2018. A total of 15 423 older adults aged 65 years and over meeting the criteria were finally included in the study. A questionnaire survey was used to collect information on demographic characteristics, lifestyle habits and self-reported prevalence of hypertension. Blood pressure values were obtained through physical examination. The level of normalized difference vegetation index(NDVI) was measured by the Medium-resolution Imaging Spectral Radiator(MODIS) of the National Aeronautics and Space Administration(NASA). The concentration of NO2 was from China's surface air pollutant data set. Meteorological data was from NASA MERRA-2. The exposure to NDVI and NO2 for each study subject was calculated based on the area within a 1 km radius around their residence. The association between mixed exposure of NDVI and NO2 as well as their interaction and hypertension in older adults was analyzed by using the multivariate logistic regression model. The restrictive cubic spline(RCS) function was used to explore the exposure-response relationship between greenness and NO2 and the risk of hypertension in study subjects. Results: The mean age of 15 423 older adults were (85.6±11.6). Women accounted for 56.3%(8 685/15 423) and 55.6%(8 578/15 423) lived in urban areas. The mean time of residence was (60.9±28.5) years. 59.8% of participants were with hypertension. The mean NDVI level was 0.41±0.13, and the mean NO2 concentration was (32.18±10.36) µg/cm3. The results of multivariate logistic regression analysis showed that NDVI was inversely and linearly associated with the hypertension in older adults, with the OR(95%CI) value of 0.959(0.928-0.992). Compared with the T1 group of NDVI, the risk of hypertension was lower in the T3 group, with the OR(95%CI) value of 0.852(0.769-0.944), and the trend test was statistically significant(P<0.05). Compared with the T1 group of NO2, the risk of hypertension was higher in the T2 and T3 groups, with OR(95%CI) values of 1.160(1.055-1.275) and 1.244(1.111-1.393), and the trend test was statistically significant (P<0.05). The result of the RCS showed that NDVI was inversely and linearly associated with hypertension in older adults. NO2 was nonlinearly associated with hypertension in older adults. The interaction analysis showed that NDVI and NO2 had a negative multiplicative interaction on the risk of hypertension, with OR(95%CI) value of 0.995(0.992-0.997). Conclusion: Exposure to greenness and NO2 are associated with hypertension in older adults.

[A panel study on the effect of atmospheric PM2.5 exposure on the gut microbiome in healthy elderly people aged 60-69 years old].

Objective: To analyze the short-term effect of individual atmospheric PM2.5 exposure on the diversity, enterotype, and community structure of gut microbiome in healthy elderly people in Jinan, Shandong province. Methods: The present panel study recruited 76 healthy elderly people aged 60-69 years old in Dianliu Street, Lixia District, Jinan, Shandong Province, and followed them up five times from September 2018 to January 2019. The relevant information was collected by questionnaire, physical examination, precise monitoring of individual PM2.5 exposure, fecal sample collection and gut microbiome 16S rDNA sequencing. The Dirichlet multinomial mixtures (DMM) model was used to analyze the enterotype. Linear mixed effect model and generalized linear mixed effect model were used to analyze the effect of PM2.5 exposure on gut microbiome α diversity indices (Shannon, Simpson, Chao1, and ACE indices), enterotype and abundance of core species. Results: Each of the 76 subjects participated in at least two follow-up visits, resulting in a total of 352 person-visits. The age of 76 subjects was (65.0±2.8) years old with BMI (25.0±2.4) kg/m2. There were 38 males accounting for 50% of the subjects. People with an educational level of primary school or below accounted for 10.5% of the 76 subjects, and those with secondary school and junior college or above accounting for 71.1% and 18.4%. The individual PM2.5 exposure concentration of 76 subjects during the study period was (58.7±53.7) µg/m3. DMM model showed that the subjects could be divided into four enterotypes, which were mainly driven by Bacteroides, Faecalibacterium, Lachnospiraceae, Prevotellaceae, and Ruminococcaceae. Linear mixed effects model showed that different lag periods of PM2.5 exposure were significantly associated with a lower gut α diversity index (FDR<0.05 after correction). Further analysis showed that PM2.5 exposure was significantly associated with changes in the abundances of Firmicutes (Megamonas, Blautia, Streptococcus, etc.) and Bacteroidetes (Alistipes) (FDR<0.05 after correction). Conclusion: Short-term PM2.5 exposure is significantly associated with a decrease in gut microbiome diversity and changes in the abundance of several species of Firmicutes and Bacteroidetes in the elderly. It is necessary to further explore the underlying mechanisms between PM2.5 exposure and the gut microbiome, so as to provide a scientific basis for promoting the intestinal health of the elderly.

[Association between urinary arsenic level and serum testosterone in Chinese men aged 18 to 79 years].

Objective: To investigate the association between the urinary arsenic level and serum total testosterone in Chinese men aged 18 to 79 years. Methods: A total of 5 048 male participants aged 18 to 79 years were recruited from the China National Human Biomonitoring (CNHBM) from 2017 to 2018. Questionnaires and physical examinations were used to collect information on demographic characteristics, lifestyle, food intake frequency and health status. Venous blood and urine samples were collected to detect the level of serum total testosterone, urinary arsenic and urinary creatinine. Participants were divided into three groups (low, middle, and high) based on the tertiles of creatinine-adjusted urinary arsenic concentration. Weighted multiple linear regression was fitted to analyze the association of urinary arsenic with serum total testosterone. Results: The weighted average age of 5 048 Chinese men was (46.72±0.40) years. Geometric mean concentration (95%CI) of urinary arsenic, creatinine-adjusted urinary arsenic and serum testosterone was 22.46 (20.08, 25.12) µg/L, 19.36 (16.92, 22.15) µg/g·Cr and 18.13 (17.42, 18.85) nmol/L, respectively. After controlling for covariates, compared with the low-level urinary arsenic group, the testosterone level of the participants in the middle-level group and the high-level group decreased gradually. The percentile ratio (95%CI) was -5.17% (-13.14%, 3.54%) and -10.33% (-15.68%, -4.63). The subgroup analysis showed that the association between the urinary arsenic level and testosterone level was more obvious in the group with BMI<24 kg/m2 group (Pinteraction=0.023). Conclusion: There is a negative association between the urinary arsenic level and serum total testosterone in Chinese men aged 18 to 79 years.

[Clinicopathological and molecular genetic features of cyclin D1-negative mantle cell lymphoma].

Objective: To investigate the clinicopathological features and molecular genetics of cyclin D1-negative mantle cell lymphoma (MCL). Methods: The clinicopathological features and molecular genetics of CyclinD1-negative MCL diagnosed between January 2016 and July 2021 at the First Affiliated Hospital of Zhengzhou University were analyzed using immunohistochemistry and fluorescence in situ hybridization. Clinical information was collected and analyzed. Results: A total of five Cyclin D1-negative MCL cases from all 212 MCL patients (5/212, 2.4%)were included. There were three male and two female patients,age ranged from 59 to 70 years (median 64 years). All patients presented with nodal lesions. None of the patients had B symptoms but four had bone marrow involvement. Histopathologically, four cases were classic MCL and one case was pleomorphic variant type. All five cases were negative for Cyclin D1 but SOX-11 were positive in all cases. CD5 was positive in four cases and one case was weakly positive for CD23. CD10 and bcl-6 were negative in all cases. CCND1 translocation was identified in three cases and CCND2 translocation in one case by FISH analysis. However,CCND3 translocations were not found in the five cases. Conclusions: Cyclin D1-negative MCL are uncommon, its accurate diagnosis needs combined analysis with morphologic and immunophenotypic characteristics and genetic changes. It may be particularly difficult to distinguish from other small cell type B cell lymphomas. FISH analyses for CCND1/CCND2/CCND3 translocations and immunohistochemistry for SOX-11 are helpful to resolve such a difficult distinction.

[Clinicopathological characteristics of natural killer cell enteropathy: report of two cases and review of literature].

Objective: To study the clinicopathological and genetic features of natural killer (NK)-cell enteropathy for better understanding of this rare disease and prevention of its misdiagnosis. Methods: Two cases of NK-cell enteropathy were diagnosed in the First Affiliated Hospital of Zhengzhou University, China from October 2017 to February 2021. The clinical characteristics, morphology, immunohistochemistry, Epstein-Barr virus-encoded RNA (EBER) in situ hybridization and T cell receptor gene rearrangement were analyzed. The patients were followed up by a telephone interview. Results: The patients were both male, aged 40 and 28 years, respectively. Both patients were admitted to the hospital for an annual checkup without obvious gastrointestinal symptoms. The endoscopy showed that the gastric body of case 1 had a mucosal bulge, small area of congestion and erosion, while the rectum of case 2 had congestion and erosion. Microscopically, the lesions of the 2 cases were relatively limited. Many lymphoid cells infiltrated within the lamina propria of the mucosa and into the muscularis mucosa in case 2. In case 1, the glands were reduced in the lesion, and the glandular cavity was slightly compressed and deformed. There was no infiltration or destruction of the glands in either case. Lymphoid cells were atypical, with medium-to-large cell sizes. Their cytoplasm was medium-to-slightly abundant and appeared eosinophilic or translucent. In case 2, characteristic eosinophilic granules were seen in the cytoplasm of a few cells. The nuclei in both cases were round, oval and irregular, with fine chromatin, inconspicuous nucleoli, and no mitotic figures were noted. Necrosis was seen in case 1 while both cases had no central growth or destruction of blood vessels. Immunophenotyping showed that CD56, granzyme B and TIA-1 were positive in both cases, part of the cells was CD3-positive, and some cells were weakly CD4-positive in case 2. The CD5, CD8, CD30, ALK and B-lineage markers (CD20, CD79α) were all negative. The Ki-67 proliferation index was about 60% and 30%, respectively. Both cases were EBER negative. TCR gene rearrangement was polyclonal. Follow-up showed that none of the 2 patients had any special treatments and stayed well. Conclusions: NK-cell enteropathy is rare, with biological behaviors similar to benign tumors, and occasional recurrence. Its histology and immunophenotype are easily confused with NK/T cell-derived lymphomas. Combination of its unique endoscopic features, EBER negativity, polyclonal TCR gene rearrangement and good prognosis can confirm the diagnosis and avoid misdiagnosis and overtreatment.

[Recommendations of diagnosis and treatment of juvenile idiopathic arthritis in China].

The common clinical subtypes of juvenile idiopathic arthritis (JIA) include systemic onset juvenile idiopathic arthritis (SOJIA), oligoarthritis/polyarthritis juvenile idiopathic arthritis and juvenile spondyloarthritis. Juvenile idiopathic arthritis has no specific diagnostic index, and needs to be differentiated from infectious diseases and malignant diseases. The onset of SOJIA is rapid, the disease progresses rapidly, and it is easy to be complicated with macrophage activation syndrome (MAS) which is life-threatening. The experience of pediatric rheumatologists in dealing with JIA is still insufficient, and the standardized diagnosis and treatment level of this disease needs to be further improved. Based on the experience and guidelines of diagnosis and treatment in China and abroad, we formulated this diagnosis and treatment standard, aiming at standardizing the diagnosis and treatment of the subtypes of JIA and MAS, so as to reduce the incidence of disability and serious complications and improve the prognosis.

[Research progress and clinical application value of exosomal circRNAs in hepatocellular carcinoma].

Primary liver cancer (hereinafter referred to as HCC) is the sixth most common type of cancer and the third leading cause of cancer-related mortality worldwide. Since patients in the early stage of HCC are typically asymptomatic and there are currently no specific detection methods for the early stage of HCC, the majority of patients are diagnosed in the late stage. Exosomes carry proteins, non-coding RNAs, such as cyclic RNAs (circRNAs), and other biological molecules. Patients with hepatocellular carcinoma have a higher concentration of serum exosomes than healthy individuals, and the circRNAs in the exosomes can reflect the source cells and real-time disease status, suggesting it has the potential to detect liver cancer early. This paper focuses on the latest progress of exosomal circRNAs and analyzes the potential value of exosomes in the early diagnosis, treatment, and progression of HCC.

[Recommendations of diagnosis of autoinflammatory diseases in China].

Autoinflammatory diseases (AIDs) are a group of disorders characterized by dysfunction of innate immunity which caused by gene mutations leading to coded proteins changes, finally causing uncontrolled systemic inflammation. AIDs are a group of rare rheumatic and inflammatory diseases. Here, Chinese Rheumatology Association summarized manifestations of the main AIDs, and to standardize the methods for diagnosis of AIDs.

[Efficacy and safety of intraoperative ultrasound-assisted neuroendoscopy in the treatment of hypertensive intracerebral hemorrhage via transsylvian approach].

Objective: To explore the efficacy and safety of intraoperative ultrasound-assisted neuroendoscopy for treating hypertensive intracerebral hemorrhage (HICH) via lateral fissure and insula approach. Methods: The clinical data of 66 patients with HICH in basal ganglia who underwent intraoperative ultrasound-assisted neuroendoscopy via lateral fissure and insula approach for hematoma evacuation were retrospectively analyzed, including operative data and follow-up results. All patients had no brain hernia before operation. The hematoma was located by the ultrasound and then the path into the hematoma cavity was determined. Meanwhile, the residual hematoma was also detected by using the ultrasound. The clearance rate of hematoma was observed by CT. Barthel index was used to evaluate the activity of daily living. Results: The amount of residual hematoma was less than 10% in 63 ases and 10%-20% in 3 cases. There were 11 cases of postoperative pneumonia, of whom 9 cases underwent percutaneous tracheotomy. Rebleeding occurred in 2 cases, but the amount was small, and there was no need for reoperation. Meanwhile, there was 1 case of secondary intracranial infection and 1 case of secondary hydrocephalus. No case of cerebral contusion was reported. Half a year after the operation, Barthel Index grading showed that there were 3 cases of grade I, 25 cases of grade Ⅱ, 34 cases of grade Ⅲ, 3 cases of grade Ⅳ and 1 case of death, respectively. Conclusions: Intraoperative ultrasound-assisted neuroendoscopy via lateral fissure and insula approach can improve the efficacy and safety of treatment for HICH.

[Screening core genes and cyclin B2 as a potential diagnosis, treatment and prognostic biomarker of hepatocellular carcinoma based on bioinformatics analysis].

Objective: To screen out and explore the core gene (Hub gene) involvement and the potential role of cyclin B2 (CCNB2) in the development and prognosis of hepatocellular carcinoma (HCC) through bioinformatics methods. Methods: Four HCC-related datasets were screened, and downloaded from the GEO database. GEO2R tool was used to analyze data and identify the differentially expressed genes (DEGs). Gene Ontology (GO) and KEGG signal pathway enrichment analysis were completed using DAVID database and Cytoscape (ClueGO) plug-in, respectively. Protein-protein interaction network (PPI) of DEGs was established using the STRING database. Cytoscape software was used to visualize PPI network, key modules (cluster) construction and core genes identification. UCSC and UALCAN database were used to analyze the differential expression and survival of TCGA hepatocellular carcinoma core genes. Firebrowse, Oncomine and UALCAN databases were used to analyze the expression of core genes in multiple tumors including HCC. Real-time quantitative reverse transcription PCR (RT-qPCR) was used to detect the expression levels of candidate genes in HCC tissues and liver cancer cell lines. Results: A total of 73 DEGs were identified from the four datasets, including 15 up-regulated genes and 58 down-regulated genes. KEGG pathway enrichment analysis signal showed that DEGs were mainly enriched in tumor-related pathways. PPI network based on DEGs had screened the key modules and 10 core genes. CCNB2 and NCAPG were highly expressed in liver cancer tissues in multiple databases. CCNB2 was positively correlated with NCAPG and was considered as a key gene related to prognosis (P < 0.01). RT-qPCR results showed that CCNB2 was highly expressed in human HCC tissues and cell lines (P < 0.01). Conclusion: Successfully screened DEGs and core genes related to HCC. Among them, CCNB2 is highly expressed in HCC and is related to the survival and prognosis of patients, so it is expected to become a biomarker for the diagnosis and prognosis of HCC.

[The prevalence and relationship between laryngopharyngeal reflux disease, anxiety and depression in otolaryngology outpatients].

Objective: To obtain the prevalence laryngopharyngeal reflux disease (LPRD), anxiety and depression in otorhinolaryngology outpatients and to explore the role of mental and psychological factors (anxiety and depression) in their pathogenesis. Methods: A questionnaire survey of reflux symptom index(RSI) scale and hospital anxiety and depression (HAD) scale were used to report 1 111 cases of outpatients in Department of Otorhinolaryngology Head and Neck Surgery, Peking University People's Hospital, from July 2017 to June 2018 (486 males, 625 females, age of 18-96 years old, median age of 38[30,53] years old) and to obtain the prevalence of LPRD, anxiety and depression. RSI-positive patients were selected in the case group, and RSI-negative patients were selected in the control group. The differences in HAD scores between the two groups were compared, and the risk factors of laryngopharyngeal reflux were analyzed. Statistical analysis was performed using SPSS 20.0 software. Results: There were 151 cases in the case group and 960 cases in the control group. The prevalence of LPRD was 13.59% (151/1 111).There was no significant difference in the prevalence of LPRD between different genders (P>0.05). The prevalence rate was the highest in the 18-40 age group, and the difference in the prevalence of all age groups (18~ 40 years old; 41-65 years old; >65 years old) was statistically significant (P<0.05). The prevalence of LPRD among smokers and non-drinkers was higher than that of non-smokers and non-drinkers and the prevalence of the two groups was statistically significant (P<0.05). The most common symptoms of the RSI scale were pharyngeal foreign body sensation (92.72%,140/151), persistent clearing throat (88.74%,124/151), excessive sputum or nasal reflux (82.12%, 124/151). There were significant statistical differences between the two groups (P<0.05). Ninty-one patients with anxiety, the prevalence was 8.19%(91/1 111); 76 patients with depression, the prevalence was 6.84%(76/1 111).Among the LPRD patients, the hospital anxiety scale scored 29.14% (44/151), and the hospital depression scale scored 17.22% (26/151). The scores of anxiety symptoms and depressive symptoms in the LPRD group were higher than those in the non-LPRD group. The above scores were statistically significant (P<0.05). Logistic regression analysis showed that smoking, anxiety and symptoms of gastroesophageal reflux disease were independent risk factors for laryngopharyngeal reflux. Conclusions: The prevalences of LPRD, anxiety and depression in the otorhinolaryngology clinic are 13.59%, 8.19% and 6.84%, respectively. Among patients with laryngopharyngeal reflux, the prevalence of anxiety is 29.14%, and the prevalence of depression is 17.22%. Age, smoking, drinking, alcohol consumption, education level, course of disease, symptoms of gastroesophageal reflux disease, pharyngeal foreign body sensation, etc. are related to LPRD. Mental factors (anxiety and depression) may play a role in LPRD. Smoking, anxiety symptoms and symptoms of gastroesophageal reflux disease are closely related to the incidence of LPRD.

[Investigation of laryngopharyngeal reflux prevalence and risk factors in otolaryngology].

Objective:To investigate the prevalence and related risk factors of laryngopharyngeal reflux in otolaryngology. Method:During January 2019 to March 2019, the inpatients in otolaryngology were investigated by the questionnaire of reflux symptom index scale and the laryngopharyngeal reflux related risk factors were analyzed. Result:Among the 227 patients, 33 patients with suspected LPR contained 19 patients（20.7%） of 92 patients in the laryngopharyngeal group, 10 patients（16.1%） of 62 patients in the nasal group, and 4 patients（5.5%） of 73 patients in the ear group. LPR prevalence in the laryngopharyngeal group was statistically different from that in the ear group（P<0.05）. Univariate logistic regression analysis showed that age, gender, smoking, drinking and BMI were risk factors of LPR（P<0.01）. Multivariate logistic regression analysis indicated that BMI was an independent risk factor of LPR（P<0.01）. Conclusion:LPR had a high prevalence rate in otolaryngology related diseases, and appropriate synergistic anti-LPR treatment could be carried out while treating otolaryngology related diseases.

[Establishment and research of a New Zealand rabbit model of laryngopharyngeal reflux].

Objective: To establish a New Zealand rabbit animal model of laryngopharyngeal reflux disease (LPRD) using esophageal balloon together with metal internal stent dilation and to investigate the changes of mucosa. Methods: 20 New Zealand rabbits were randomly divided into experimental group and control group, with 10 in each group. Balloon dilatation and metal internal stent dilation were carried out in experimental group to reproduce the animal model of LPRD.The middle of balloon was placed at the lower esophageal sphincter (LES) while the stent was placed at the upper esophageal sphincter (UES). The guide wire was placed in the control group, but the balloon was not expanded and the stent was not placed. The general condition, pH value of hypopharynx, laryngeal histopathology and changes of pepsin content of New Zealand rabbits were observed regularly. The difference between experimental group and control group was compared. Results: The 24-hour Dx-pH monitoring results showed that the number of reflux episodes(20.0[9.5, 35.0], 13.0[6.5, 22.0]), and the percent time below pH 5.5 (1.36%[0.60%, 4.57%], 1.36%[0.43%, 2.77%]) in the experimental group at the 2nd and 4th week were significantly different from those in the control group (0[0,3.0], 1.0[0.5, 3.8]; 0[0, 0.01%], 0[0, 0], respectively, all P<0.01), suggesting that the experimental group New Zealand rabbits developed LPRD. Compared with the control group under microscope, lymphocytes infiltration and submucosal gland hyperplasia increased in the mucosa of the throat of the experimental group. The results of pepsin immunohistochemical staining between the two groups were statistically significant (P=0.014). Conclusion: The use of balloon dilatation of the LES combined with metal stent dilatation of the UES can successfully establish a laryngopharyngeal reflux model, and lesions in the throat tissue can be observed.

[Effect and mechanism of adipocyte co-culture on aquaporin-9 expression in HepG2 cells].

Objective: To observe the effect of differentiated mature adipocytes on hepatic steatosis and aquaporin-9 (AQP9) expressions in HepG2 cells and further explore its possible mechanism of action. Methods: Human preadipocytes were cultured and differentiated to full maturity. HepG2 cells were co-cultured with non-differentiated adipocytes and differentiated mature adipocytes for 48 h, and then labeled as control group and experimental group. Oil red O staining and intracellular triglyceride content were performed on co-cultured HepG2 cells and simultaneous changes in phosphatidylinositol 3-kinase (PI3K) - serine/threonine kinase (Akt) signaling pathway, and AQP9 mRNA and protein levels were detected. The experimental group was co-cultured with recombinant human insulin-like growth factor-I (IGF-I), with the addition of 100ng/ml PI3K-Akt pathway agonist, labeled as experimental group + IGF-I group. The activation of PI3K-Akt pathway was verified by Western blotting (WB). The expression of AQP9 was detected by RT-q PCR and WB. The recombinant lentivirus LV-AQP9 or empty-loaded virus LV-PWPI was transfected with HepG2 cells by recombinant lentiviral transfection tecnique, and labeled as HepG2-AQP9 and HepG2-PWPI. The transfection efficiency was assessed by confocal laser scanning microscopy and RT-qPCR and WB detected the change of AQP9 expression level after virus transfection. Afterwards, the stable over-expressed HepG2-AQP9 cells and the empty-loaded HepG2-PWPI cells were co-cultured with differentiated mature adipocytes for 48h, and labeled as HepG2-AQP9 co-culture group, and then intracellular triglyceride content were detected with Oil red O staining. Finally, IGF-I was added to the HepG2-AQP9 co-culture group, which was recorded as HepG2-AQP9 co-culture + IGF-I group. Intracellular triglyceride content was detected with Oil red O staining, and WB verified PI3K-Akt signaling pathway activation and changes in AQP9 mRNA and protein levels. A t-test was used to compare the two independent samples. Results: The intracellular lipid droplets and triglyceride content (0.052 ± 0.005) in the experimental group was increased significantly than the control group (0.033 ± 0.003) (t= 5.225,P= 0.006), suggesting that adipocyte co-culture had induced steatosis in HepG2 cells. RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (3.615 ± 0.330) and protein levels (0.072 ± 0.005) in the experimental group were significantly higher than the control group (t= 13.708, 11.225,P= 0.005, < 0.001). WB results showed that the expression level of phosphorylated Akt (p-Akt) protein (0.116±0.003) in the experimental group was significantly lower than the control group (0.202 ± 0.003) (t= 27.136,P< 0.001). The total Akt protein was constant, and the p-Akt/total Akt (0.182 ± 0.017)was significantly lower than the control group (0.327 ± 0.019) (t= 2.431,P= 0.001), suggesting that adipocyte co-culture had inhibited PI3K- Akt signaling pathway in HepG2 cells and up-regulated the expression level of AQP9. WB results indicated that the expression level of p-Akt protein (0.194 ± 0.021) in the experimental group + IGF-I group was significantly higher than the experimental group (0.132 ± 0.003) (t= 5.082,P= 0.007). The total Akt protein was constant, and the p-Akt/total Akt (0.281 ± 0.009) was significantly higher than the control group (0.184 ± 0.132) (t= 10.311,P< 0.001). Simultaneously, RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (0.327 ± 0.347) and protein levels (0.042 ± 0.004) in the experimental group + IGF-I group were significantly lower than the experimental group (t= 33.573, 5.598,P< 0.001, 0.005), suggesting that adipocyte co-culture had possibility to regulate the expression level of AQP9 through the PI3K-Akt pathway. Confocal laser microscopy analysis showed that the transfection efficiency was more than 90%. RT-q PCR and WB results indicated that the expression levels of AQP9 mRNA and protein levels (0.373 ± 0.221) in HepG2-AQP9 group were significantly higher than HepG2-PWPI group (t=14.953, 28.931,P= 0.002 and 0.000), suggesting that the stable overexpression of AQP9 cell line was successfully constructed. The intracellular lipid droplets and triglyceride content in HepG2-AQP9 co-culture group was significantly increased (t= 5.478, 5.369,P= 0.005) than HepG2-PWPI co-culture group and HepG2-AQP9 co-culture+ IGF-I group, suggesting that the increased expression of AQP9 had promoted HepG2 steatosis in co-cultured adipocytes. WB results showed the expression levels of p-Akt protein (0.168 ± 0.006) and p-Akt/total Akt (0.265±0.009) in HepG2-AQP9 co-culture + IGF-1 group was significantly increased (t= 16.311, 8.769,P< 0.001) than HepG2-AQP9 co-culture group, while the expression levels of AQP9 mRNA (0.327 ± 0.034) and protein (0.375 ± 0.025) was significantly decreased (t= 33.573, 9.146,P< 0.001 and 0.001). Conclusion: Adipocytes co-culture can induce steatosis in HepG2 cells, and may participate in inhibiting PI3K-Akt signaling pathway to upregulate the expression of AQP9 in steatotic HepG2 cells.

[Correlation between CTL and Th1 cell proportion in peripheral blood of liver transplantation recipients and the success of hepatitis B vaccination].

Objective: To investigate the correlation between the proportion of CTL and Th1 cells in peripheral blood of liver transplant recipients and the success of hepatitis B vaccination. Methods: The subjects of this study were liver transplantation recipients with chronic HBV-related liver diseases in Organ transplantation institute of the third medical center of PLA general hospital. Subjects were randomly divided into two groups for prospective study. In the rapid group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2and 3 months, and one dose of 20 µg hepatitis B vaccine was administered at 4, 5 and 6 months. In the rapid-enhanced group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2 and 3 months, and one dose of 60 µg hepatitis B vaccine was administered at 4, 5 and 6months. Compare and analyze the success rate of inoculation, the titer of hepatitis B surface antibody (anti-HBs), the proportion of CTL cells in CD8(+)T cells and Th1 cells in CD4(+)T cells. Correlation analysis was performed for CTL and Th1 cells and anti-HBs, Observe the safety of vaccination. Results: The inoculation success rate, anti-HBs growth rate, CTL cell percentage increase and Th1 cell percentage increase in the rapid enhancement group were all higher than those in the rapid enhancement group, and the differences were statistically significant, they were 38.3% (23/60) vs 21.7% (13/60) (P=0.046), 91.3(72.5,124.2) vs 22.1(12.4, 31.6) (P=0.001), 1.4(0.8,1.9) vs 0.4(0.2,1.4) (P=0.001) and 7.4±2.6 vs 5.6±3.7 (P=0.001) respectively. The percentage increase of CTL cells and Th1 cells in the successful group was greater than that in the non-successful group, and the difference was statistically significant. They were 1.9(1.4,2.5) vs 0.1(0.0,1.1) (P=0.024) and 9.6±3.1 vs 2.4±2.0 (P<0.001). There was no significant correlation between anti-HBs increase (105.5±37.1) and CTL increase 1(0,3) (P=0.099), while there was significant positive correlation with Th1 increase 7(2,11) (P<0.001). No rejection reaction occurred during the study period, and there was no special abnormal change in the safety index. Conclusion: Reasonable increase of vaccine dose can up-regulate Th1 cell expression and promote the generation of anti-HBs.

[Analyses of pathogenic factors and clinicopathological characteristics of vocal leukoplakia].

Objective: To investigate the pathogenic factors of vocal leukoplakia and its clinical and pathological features. Methods: Eighty-one patients with vocal cord leukoplakia who underwent surgery between February 2010 and December 2016 and 160 volunteers without pharyngeal symptoms designed as controls were included in this case control study. The clinicopathological characteristics of 81 patients were summarized and analyzed synthetically. Results: There was statistical significance in reflux symptom index(RSI), reflux finding score(RFS), smoking index, and drinking index between case group and control group(Z=-5.35, -4.82, -4.76, -2.44, P<0.05). The voice-using duration per day in case group was significantly longer than that of control group.There was no statistical significance in hospital anxiety and depression scale for anxiety(HADA) scores、hospital anxiety and depression scale for depression(HADD) scores between case group and control group(P>0.05). In 42 patients who received 24-hour dual probe pH monitoring the prevalence of pathologic LPR was 42.8%. In 81 patients, 39(48%)patients were pathologically diagnosed as squamous cell hyperplasia, 18(22%)patients as mild dysplasia, 12(15%)sides as moderate dysplasia , 10(12%)patients as severe dysplasia and 2(2%)patients as carcinoma in-situ. The average age of high-risk pathological vocal leukoplakia was significantly higher than that of low-risk leukoplakia(t=-2.73, P<0.01). The propotion of speckled leukoplakia in high-risk leukoplakia was significantly higher than that of low-risk leukoplakia(χ(2)=23.81, P<0.01). There was no statistical significance between high-risk leukoplakia and low-risk leukoplakia in the prevalence of pathologic LPR(P>0.05). The bilateral lesions, speckled leukoplakia were more likely to relapse(χ(2)=4.27, 12.17, P<0.05). The more serious the pathology, the more likely it was to relapse (Z=-2.168, P=0.03). There was no statistical significance between recurrence group and non-recurrence group in the prevalence of pathologic LPR(P>0.05). Conclusions: LPR, smoke constitute the risk factors of vocal cord leukoplakia. Drinking, voice abuse are related to vocal cord leukoplakia. Senile, speckled leukoplakia are more likely to be malignancy. A speckled leukoplakia, bilateral leukoplakia, severe pathological degree are important factors to predict recurrence.

[Analysis on 10 year survival of HIV/AIDS patients receiving antiretroviral therapy during 2003-2005 in Henan province].

Objective: To understand the survival of HIV/AIDS patients after receiving antiretroviral therapy for 10 year in Henan province and related factors. Methods: The database of national integrated management system of HIV/AIDS was used to collect the basic information and follow-up information of HIV/AIDS patients who received antiretroviral therapy between 2003 and 2005 in Henan province. Software SPSS 23.0 was used to analyze the patients' survival and related factors based on the life-table method and Cox proportional hazards model. Results: Among the 2 448 HIV/AIDS patients who started antiretroviral therapy during 2003-2005, the men accounted for 53.5%, and women accounted for 46.5%. Up to 70.1% of the patients were aged 40-59 years and 95.5% of the patients had blood borne infections. The patients were observed for 10 years after antiviral treatment, and 719 cases died from AIDS related diseases, with a mortality rate of 3.78/100 per year (719/19 010 per year). The cumulative survival rates of patients within 1-year, 3 years, 5 years and 10 years were 0.94, 0.86, 0.78, 0.69 respectively. Compared with the patients aged <40 years, the HRs of the patients aged 40-, 50-, 60- and ≥70 years were 1.417 (95%CI: 0.903-2.222), 1.834 (95%CI: 1.174-2.866), 2.422 (95%CI: 1.539-3.810) and 3.424 (95%CI: 2.053-5.709) respectively. Compared with patients with baseline CD(4+)T lymphocyte >350 unit/ul, the HRs of the patients with CD(4+)T lymphocyte <50 unit/µl, 50-199 unit/ul and 200-350 unit/ul were 7.105 (95%CI: 5.449-9.264), 4.175 (95%CI: 3.249-5.366) and 2.214 (95%CI: 1.691-2.900) respectively. Compared with the women, the HR of the men was 1.480 (95%CI: 1.273-1.172). Compared with the patients who received second line ART therapy, the HR of patients receiving no second line therapy was 11.923 (95%CI: 9.410-15.104). Conclusions: The cumulative survival rate the HIV/AIDS patients after 10 years of antiretroviral therapy reached 0.69 in Henan. Male, old age, low basic CD(4+)T lymphocyte count and receiving no second line therapy were the risk factors for long-term survival of AIDS patients.