Objective To investigate the effect of hepatitis B virus X protein (HBx) on the induction of eytochrome P450 (CYP) 3A4 by 1α, 25-(OH)2D3 in HepG2 cells in vitro. Methods HepG2 cells were transiently transfected with plasmid pEGFP-N1 (control) or co-transfected with recombinant HBx eukaryotic expression plasmid pcDNA3-X and pEGFP-N1. All HepG2 cells were divided into four groups: control group (without transfection), plasmid pEGFP-N1 transfection group, plasmid pEGFP-N1 transfection plus 1α ,25-(OH)2D3 group, plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α ,25-(OH)2D3 group. The expression of CYP3A4 in HepG2 cell was induced by 0.35 μ mol/L 1α ,25-(OH)2D3 for 72 h, and mRNA levels and protein levels of CYP3A4 in the cells were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and Western-blot assay, respectively. The comparison between groups was done by F test. Results CYP3A4 mRNA level in plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α ,25-(OH)2D3 group was 1.52 folds of control group, while that in plasmid pEGFP-N1 transfection plus 1α, 25-(OH)2D3 group was 3.97 folds (F= 4.72, P<0. 05). Similarly, intracellular CYP3A4 protein expression in plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α , 25-(OH)2D3 group increased to 2.1 folds of control group, while that in plasmid pEGFP-N1 transfection plus 1α,25-(OH)2D3 group increased to 5.9 folds (F=4.68, P<0.05). Conclusion HBx interferes with the induction of CYP3A4 by 1α , 25-(OH)2D3 in HepG2 cell line, which suggests that HBx has suppressive effect on the expression of CYP3A4.

Objective: To study the chemical constituents of the methanol extract from the aerial parts of the mangrove plant Sonneratia paracaseolaris. Methods: The methanol extract was isolated and purified with various chromatographic methods, including silica gel, ODS, Sephadex LH-20 columns, TLC, and HPLC. The compounds were identified by their physical chemical properties and 1H-NMR and 13C-NMR data. Results: Seventeen compounds were obtained from the methanol extract of Sonneratia paracaseolaris and identified as phytol (1), stigmasta-4-ene-3,6-dione (2), stigmata-4,22-diene-3,6-dione (3), cholesterol (4), (22E)-cholesterol-5,22-diene-3β-alcohol (5), diosmetin (6), tricin (7), 5,3′,5′-trihydroxy-7,4′-dimethoxyflavone (8), 5-hydroxy- 7,4′-dimethoxyflavone (9), 5-hydroxyl-7,3′,4′-trimethoxydihydroflavone (10), vanillin (11), p-hydroxy benzaldehyde (12), salicylic acid (13), trans-p-hydroxyl ethyl cinnamate (14), 4-hydroxy-2,6-dimethoxy-benzaldehyde (15), 3,4,5-trimethoxybenzoic acid (16), and 3,3′,4-trimethoxyellagic acid (17). Conclusion: All the compounds except 4, 11, 15, and 17 are obtained from genus Sonneratia. All compounds are isolated from S. paracaseolaris for the first time.

OBJECTIVETo establish a convenient and efficient model for investigating the expression of CYP3A4 and drug metabolism in vitro.

METHODS1alpha,25-dihydroxyvitamin D3 was utilized as an inducer to enhance CYP3A4 expression in HepG2 cells. 0.1, 0.25, 0.35 micromol/L 1alpha,25-dihydroxyvitamin D3 were added to the cell culture media, and cells were harvested after 24, 48, 72 and 96 hours. Cell proliferation was determined with MTT assay. CYP3A4 mRNA level was analyzed with RT-PCR and expressions of CYP3A4 protein were measured by Western blot.

RESULTS1alpha,25-dihydroxyvitamin D3 in 3 concentrations, namely 0.10, 0.25 and 0.35 micromol/L, did not show obvious toxicity to HepG2 cells. At 24 h of the cultivation, the expression of CYP3A4 mRNA was not increased significantly, but CYP3A4 mRNA expression significantly increased by 120%, 134%, 200% at 48 h, by 174%, 254%, 420% at 72 h, and by 258%, 450%, 370% at 96 h, respectively under the three concentrations. Similar results were observed in the induction of CYP3A4 protein expression. At 48, 72 and 96 hours after treatment with 0.25 micromol/L and 0.35 micromol/L 1alpha,25-dihydroxyvitamin D3, CYP3A4 protein increased in various folds in the controls (1.2 and 2.2 after 48 h, 3.4 and 6.5 after 72 h, 6.1 and 7.2 after 96 h), while 0.10 micromol/L 1alpha,25-dihydroxyvitamin D3 only induced protein expression at 72 h and 96 h (1.8 and 4.1 folds, respectively).

CONCLUSION1alpha,25-dihydroxyvitamin D3 could induce the expression of CYP3A4 mRNA as well as CYP3A4 protein in HepG2, which provides a convenient and efficient in vitro system for investigation of CYP3A4 and drug interaction.

Calcitriol ; pharmacology ; Cytochrome P-450 CYP3A ; drug effects ; genetics ; metabolism ; Hep G2 Cells ; Humans ; Transcription, Genetic

[ ABSTRACT] The splicing factors were characterized for their crucial roles in pre-mRNA splicing of eukaryons. SRSF2 is a member of the SR protein family which is one of the most common splicing factors, and it is believed to be a key element in pre-mRNA splicing, mRNA transcription, regulation of the DNA stability and cell proliferation.SRSF2 gene mutation is detected frequently in myeloid malignancies ( like MDS and CMML) and may be associated with the phenotype and prognosis of these malignancies.The paper makes a review for the latest research progression on SRSF2 gene mutation and its relationship with myeloid malignancies.

Objective Idiopathic pulmonary fibrosis ( IPF) is a chronic inflammatory disease with unknown etiology and is lack of effective therapy. The aim of this study is to explore the function of Ca2+ and PI3K/AKT/mTOR pathway in the pathogenesis of IPF, and the impact of 1,25?( OH) 2 D3 on Ca2+ and PI3K/AKT/mTOR pathway in type Ⅱalveolar epithelial cells of rat with IPF. Methods 150 male SD rats were randomly divided into 2 groups: prevention group ( control groupⅠ, model groupⅠ, medication groupⅠ) and treatment group ( control groupⅡ, model groupⅡ, medication groupⅡ) . The tracheal exposure surgery was operated in control groupⅠ/Ⅱ, and then 200μL sterile physiological saline was administered by intraperitoneal injection of each rats 2 days and 14 days after surgery, separately. Bleomycin(BLM)(5 mg/kg) was in?jected into the trachea of model groupⅠ/Ⅱ, and then vitamin D3 solvent(0.1%ethanol and 99.9%glycol propylene, 1μL/g) was ad?ministered by intraperitoneal injection 2 days and 14 days after surger?y, separately. Bleomycin( BLM) ( 5 mg/kg) was injected into the tra?chea of medication groupⅠ/Ⅱ, and then 1,25?( OH) 2 D3( 2μg/kg) was administered by intraperitoneal injection 2 days and 14 days after surgery, separately. IPF model was built by injecting Bleomycin into the trachea of rats, 1,25?(OH)2D3(2μg/kg) was used to prevent and treat IPF by intraperitoneal injection in medication group. The hydroxyproline content of lung tissue in each group was measured, type Ⅱalveolar epithelial cells were separated from lung tissue and labeled with Fluo?3AM, then concentration of Ca2+ was detected by Laser scanning confocal microscope. The mRNA levels of PI3K, AKT and mTOR in the typeⅡalveolar epithelial cells were tested by RT?PCR. Results Compared with control groupⅠ/Ⅱ at each time point, hydroxyproline content of lung tissue, Ca2+ concentration and expression of PI3K, AKT and mTOR in typeⅡalveolar epithelial cells in model groupⅠ/Ⅱand medication groupⅠ/Ⅱwere sig?nificantly raised( P<0.05 or P<0.01) , but these were significantly reduced in medication groupⅠ/Ⅱcompared with model groupⅠ/Ⅱ( P<0.05 or P<0.01) . Correlation analysis showed that there is significant positive correlation between Ca2+ concentration and mRNA expression levels of PI3K, AKT and mTOR in model groupⅠ/Ⅱ(r=0.5988, r=0.6230, r=0.6603,P<0.01)and medication groupⅠ/Ⅱ( r=0.701 2, r=0.632 3,r=0.740 3,P<0.01) . Conclusion The PI3K/AKT/mTOR pathway plays an important role in devel?opment of IPF. 1,25?( OH) 2 D3 is able to reduce Ca2+concentration in typeⅡalveolar epithelial cells and inhibit the PI3K/AKT/mTOR pathway, and then inhibit the development of IPF.

Objective Not much information is available on the comparative analysis of G test on diagnosis of deep fungal in -fection by colormetric and turbidimetric measurements .The purpose of this paper was to explore the clinical value of fungal (1-3)-β-D-glucan detection kit ( colormetric measurement ) . Methods 89 clinical samples collected from Hainan Branch of PLA General Hos-pital were detected by fungal (1-3)-β-D-glucan detection kit (turbidimetric measurement) and fungal (1-3)-β-D-glucan detection kit ( colormetric measurement ) respectively , among which 32 cases were from disease group ( deep fungal infection in patients ) and 57 ca-ses were from control group ( healthy person ) .The comparison was made on the sensitivity , specificity and accuracy of these two meth-ods. Results The sensitivity, specificity and accuracy of colormetric measurement kit on the diagnosis of deep fungal infection were obviously higher than those of turbidimetry method kit (81.2%vs 53.1%, 91.2%vs 75.4%, 87.6%vs 67.4%, P<0.05), which was of significant difference .Detection result of colormetric method had a positive coincidence rate with other systems '( except respira-tory system) deep fungal infection, which was obviously higher than turbidimetry method (92.8%vs 57.1%, P<0.05).Positive co-incidence rate and total coincidence rate between colormetric method kit and clinical diagnosis result on differentiate samples were obviously higher than those of turbidimetric method kit (P=0.01). Conclusion Colormetric measurement kit has higher accuracy and higher coincidence rate with clinical diagnosis than turbidimetry meas-urement kit , which is better for clinical service .

Abstract: Objective　To explore the relationship between body mass index (BMI), waist-hip ratio (WHR) and the risk of gout in Urumqi. Methods　A total of 516 male patients with gout in a third-class hospital in Urumqi from 2015 to 2019 were randomly selected as the gout group and 516 male healthy subjects in the same hospital as the control group. The relevant blood biochemical indexes were examined and analyzed. Blood pressure, waist circumference and hip circumference were measured. Body mass index and waist-to-hip ratio were calculated. Logistic regression model was used to analyze the relationship between overweight / obesity, waist-to-hip ratio and the risk of gout. The test level is α = 0.05. Results　Uric acid, glucose, urea nitrogen, creatinine, triglyceride, low-density lipoprotein, systolic blood pressure, weight and waist circumference in gout group were higher than those in control group, and the differences were statistically significant (P<0.05); There were no significant differences in age, height and diastolic blood pressure between the two groups (P<0.05). There was a positive correlation between BMI and WHR and the occurrence of gout (r=0.272, 0.345, P<0.05). There were significant differences in BMI, WHR and waist circumference between the gout group and the control group（χ2= 55.338, 54.928, 54.153, P<0.05）. After adjusting for age, aerobic exercise and other confounding factors, the results of multi-factor unconditional Logistic regression analysis showed that the odds ratio (OR) of gout in patients with BMI of 24.00-27.99 kg/m2 and ≥28.00 kg/m2 was 2.005 (1.337-3.006) and 2.677 (1.668-4.296) times higher than that of patients with normal BMI, respectively. The OR value of gout in patients with WHR≥0.90 was 1.668 times higher than that in patients with normal WHR, and the difference was statistically significant. The results of subgroup analysis according to age are generally similar. Conclusions　The BMI and WHR of man with gout in Urumqi are higher than those of normal people, and BMI, waist circumference and WHR are all associated with the incidence of gout. The risk of gout increases with the increase of BMI and WHR.

Objective: To analyze the epidemiological characteristics of HIV-infected pregnant women and exposed infant in Guangdong province and identify the factors associated with infant HIV infection through mother-to-child transmission. Methods: National Information System for Prevention of mother-to-child HIV Transmission and Early Infant Diagnosis Information Management Platform were used to collect the individual information about HIV-infected pregnant women and exposed infants who were delivered in Guangdong from January 1, 2014 to December 31 in 2017. The differences in pregnant women’s demographic data, history of pregnancy and childbirth, the utilization of mother-to-child transmission prevention services and early infant diagnosis between the infected HIV exposed infants and uninfected HIV exposed infants were compared, and univariate and multivariate logistic regression analyses were conducted to identify the factors associated with mother-to-child HIV transmission. Results: Among 349 HIV infected pregnant women, the proportions of the pregnant women whose HIV infection status were confirmed before pregnancy, during pregnancy and at or after childbirth were 30.4% (106/349), 49.6% (173/349) and 20.0% (70/349) respectively. The proportions of those with sexual partners whose HIV infection status were unknown and those receiving no antiviral treatment were 39.5% (138/349) and 13.2% (46/349) respectively. Among the HIV exposed infants, the mother-to-child transmission rate was 4.2%(15/353), the HIV exposed infants had the first or second early diagnosis tests within 44 (P₂₅-P₇₅: 42-50) days and 96 (P₂₅-P₇₅: 92-106) days after birth, respectively. Univariate logistic regression analysis indicated that the risk for mother-to-child HIV transmission increased in those whose HIV infection status were confirmed at or after childbirth compared with before pregnancy (OR=5.72, 95%CI: 1.52-21.61) and in the group that antiviral treatment was given to either mothers or infants compared with the group that antiviral treatment was given to both mothers and infants (OR=33.56, 95%CI: 9.04-124.55), while there was lower mother-to-child HIV transmission risk in artificial feeding group compared with breast feeding group (OR=0.07, 95%CI: 0.01-0.76). Conclusion The risk of mother-to-child HIV transmission in Guangdong can be effectively reduced by the measures of early diagnosis, antiviral treatment and artificial feeding as well as the improvement of mother-to-child transmission prevention service.

Objective To analyze the differences in the expression levels of the lncRNA MALAT1, NEAT, NEAT2 in peripheral blood mononuclear cell (PBMC) from tuberculosis patients and healthy controls. Methods We detected the lncRNA expression levels in PBMC from 79 tuberculosis patients and 82 healthy controls by quantitative reverse transcription polymerase chain reaction, and analyzed the correlation between lncRNA expression levels and some clinical features and laboratory indicators in tuberculosis patients. Results The expression levels of MALAT1, NEAT1 in PBMC of tuberculosis patients were significantly higher than healthy controls (Z=-4.386, P<0.001; Z=-10.175, P<0.001). There was no significant difference in the expression of NEAT2 between tuberculosis patients and healthy controls (Z=-0.203，P=0.839). The correlation results of lncRNA levels and some clinical features, laboratory indicators in tuberculosis patients suggested that the NEAT2 level in PBMC of newly treated tuberculosis patients was higher than recurrent tuberculosis patients, while the NEAT2 level in PBMC of sputum smear positive tuberculosis patients was lower than that of sputum smear negative tuberculosis patients (all P<0.05). There was a negative correlation between MALAT1 level and erythrocyte sedimentation rate (rs=-0.256, P=0.034). Conclusion MALAT1 and NEAT1 are abnormally expressed in PBMC of tuberculosis patients, and may be involved in the pathogenesis of pulmonary tuberculosis.

OBJECTIVETo investigate the effect of penehyclidine hydrochloride (PHC) in a rat model of renal injury induced by hemorrhagic shock and lipopolysaccharides (LPS).

METHODSForty-five healthy Wistar rats were randomized into sham operated group, model group, and 3 penehyclidine hydrochloride (PHC) dose (1, 2 and 3 mg/kg) groups (PHC1, PHC2, and PHC3 groups, respectively). The arterial blood samples were collected to determine the concentrations of serum tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-1 (IL-1), urine creatinine (Cr) and blood urine nitrogen (BUN), and the renal tissues were collected to measure the expressions of ICAM-1 and nuclear factor-κB (NF-κB) and observe the pathological changes.

RESULTSTNF-α, IL-8, IL-1, Cr, BUN, ICAM-1 and NF-κB in the 3 PHC groups were significantly lower than those in the model group (P<0.05). TNF-α, IL-8, IL-1, Cr and BUN were significantly lower in PHC1 (P<0.05) than in the PHC2 and PHC3 groups, and ICAM-1 and NF-κB were similar between 3 PHC groups (P>0.05). Compared with the model group, the 3 PHC groups showed lessened pathological changes in the renal tubules.

CONCLUSIONPHC has protective effects against renal injury induced by hemorrhagic-endotoxin shock in rats, and treatment with 1 mg/kg PHC produces the most significant protective effect.

Acute Kidney Injury ; drug therapy ; etiology ; Animals ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1 ; blood ; Interleukin-8 ; blood ; Kidney ; drug effects ; metabolism ; Kidney Tubules ; drug effects ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; NF-kappa B ; metabolism ; Quinuclidines ; pharmacology ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; blood ; metabolism ; Tumor Necrosis Factor-alpha ; blood