Objectives To explore the correlation of level of serum 25-hydroxyvitamin D with pulmonary function in adult with asthma. Methods Patients were divided into Asthmatic group(n=62)and Control group (n=28). The Asthmatic group was further divided into Mild Group (n=6), Moderate Group (n=13) and Severe Group (n=43). Serum levels of 25-hy?droxyvitamin D [25(OH)D], denoted as 25(OH) Vit D was detected by ELISA. Pulmonary function indicators,including FVC (forced vital capacity) , FEV1 (forced expiratory volume in 1 second) , FEV1%predicted , and FEV1/FVC%were deter?mined by a pulmonary function testing device. General profiles such as medical history, age and height as well as serum VitD levels were compared between subgroups of the asthmatic groups and between two genders. Serum levels of 25 (OH) VitD were compared between asthmatic group and control group while its correlation with FEV1%predicted were calculated in all three sub asthmatic groups. Results There was no significant difference in medical history, age, height and the 25(OH) VitD levels between male and female participants. Serum 25(OH) Vit D level was significantly lower in the asthmatic patient group [(29.69±20.45) nmol/L] compared to that in control group [(75.16±4.06) nmol/L] (P<0.05). It was significantly lower in severe sub group than those in the mild and moderate sub groups. The differences were both statistically significant ( P<0.05). There were positive correlations between serum 25(OH) Vit D levels and FEV1%predicted ( P<0.05) in all sub asth?matic groups. Conclusion Vitamin D deficiency is highly prevalent in asthmatic patients, and there is a strong correlation between 25(OH) Vit D asthma severity as well as between lung function.

Background and purpose:New treatment strategies should be explored for non-small cell lung cancer (NSCLC) patients after the failure of the epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI). To compare the efficacy and toxicities of chemotherapy in combination with EGFR-TKI or single chemotherapy in advanced NSCLC patients with EGFR-TKI resistence. Methods:In this study, 18 patients were enrolled. Eight patients were treated by chemotherapy combined with EGFR-TKI (CE group);10 patients were treated by single chemotherapy (E group), 21 days for one cycle. All patients received at least 2 cycles of treatment. Results:All 18 patients had been evaluated. The CE group was similar to the E group in objective response rate (ORR:25%vs 10%, P=0.832). The CE group was higher than the E group in disease control rate (DCR:87.5%vs 30%, P=0.046). The median PFS was longer in CE group (3.5 months vs 2.4 months, P=0.05). The CE group was higher than the E group in rash (75%vs 10%, P<0.05). The grade 3-4 toxicities did not have significantly differences between the two groups (P>0.05). Conclusion:Though there was no significant difference in ORR between the 2 groups (P>0.05), the CE group was superior to the E group in DCR and PFS. Patients with retreatment of advanced NSCLC after the failure of EGFR-TKI can be controlled by continued EGFR-TKI and chemotherapy.

OBJECTIVE To explore the safe and effective dose of sirolimus (Rapamycin,Sir) and its effect on seizure comorbidities. METHODS Immature C57BL/6 mice at postnatal 10 d of age were administered with kainic acid(KA) 12.0 mg · kg-1 intraperitoneally by a single injection to induce acute seizure. Sir 0.3, 1.0 and 3.0 mg · kg-1 was injected 24 h after seizure every other day until 3 d, 1 week, 3 weeks, 5 weeks and 6 weeks. Western blotting analysis was used to detect the expression and phos?phorylation level of S6 protein and to determine the minimum effective dose of Sir. Effect of the mini?mum effective dose of Sir on cognitive function and body growth was observed by several evaluations. Immunofluorescent intensity of Doublecortin (DCX) immunofluorescent staining was conducted to evaluate the development of neurons in the hippocampus. Morris water maze was used to assess the cognitive function. Tail suspension test, O maze and new object recognition test were used to study the anxiety-like behaviors of mice. RESULTS The result of Western blotting showed that Sir 0.3 mg · kg-1 had no significant effect on the phosphorylation of S6 protein in normal mice or KA mice, whereas 1.0 and 3.0 mg · kg- 1 could significantly inhibit the phosphorylation of S6 protein in KA mice (P<0.05). Sir 1.0 mg·kg-1 had no obvious effect on DCX-positive cells or body wass. Morris water maze showed that KA-induced seizure resulted in prolonged escape latency and swimming length (P<0.05), and a decreased crossing number of target quadrant (P<0.05). Sir 1.0 mg·kg-1 significantly reversed the deficit of cognitive function of KA-induced seizure mice (P<0.05), whereas no significant difference was found between Sir group and normal control group. Compared with normal control group, model group showed increased freezing time in tail suspension test (P<0.05), decreased migration length and reten?tion time in open arms in O maze (P<0.05), decreased retention time and touch frequency with new objects, migration length and average speed in new object recognition test (P<0.05). Sir 1.0 mg · kg-1 significantly reversed the above anxiety and depression status, whereas no significant difference was found between sirolimus group and normal control group. CONCLUSION Sir 1.0 mg · kg-1 inhibits the abnormal activation of mTOR pathway and the formation of epilepsy comorbidity in immature mice. Along with its mild side effect in development, Sir 1.0 mg · kg-1 will be an ideal dose to be used in the treatment of seizure in immature mice.

An analytical method of fluorescent dye-labeled oligonucleotides was established by ion pair reversed phase high performance liquid chromatography(IP-RP-HPLC) which was improved by optimizing the effects of triethylamine-acetic acid(TEAA)(0-0.15 mol/L), pH(4.5-7.0) and gradient. Comparing the retention of 5, 10 and 15-mer unlabeled oligonucleotides with that of 5'-carboxyfluorescein(5'FAM) labeled oligonucleotides, the mechanism of fluorescent dye-labeled oligonucleotides retention was studied. In addition, TaqMan~(TM) probes as wellas other common fluorescent dye-labeled oligonucleotides were concerned. The results showed that the best resolution of different length fluorescent dye-labeled oligonucleotides was observed under the condition of 0.01 mol/L TEAA and pH 7.0. The retention behavior of fluorescent dye-labeled oligonucleotides was different from that of unlabeled oligonucleotides significantly, and therefore they can be separated completely. The results indicated that the retention of unlabeled oligonucleotides enhanced with the increase of the length of molecule. In contrast, the retention of fluorescent dye-labeled oligonucleotides was reduced with the increase of the length of molecule. For the hydrophobicity of fluorescent dyes made a great impact on the retention, a longer retention time the labeled oligonucleotides would take while the hydrophobicity of fluorescent dyes was higher. However, the effect of the hydrophobicity was limited as the length was increased to a certain level.

OBJECTIVE:To study the effects of paeonol on Wnt and BMP/Smad pathway in ankylosing spondylitis (AS) model mice,and to investigate prevention and treatment mechanism of paeonol for AS. METHODS:40 mice were randomly divided into normal group,model group,sulfasalazine group(positive control,9 mg/kg)and paeonol group(3 mg/kg),with 10 mice in each group. In addition to the normal group,Freund's adjuvant and proteoglycan were used to establish AS model in each group. After modeling,mice in each administration group were given relevant medicine intragastrically;normal group and model group were given constant volume of distilled water intragastrically,once a day,for consecutive 20 d. After last administration, the mice were killed,TEM was used to observe the ultrastructural pathologic change of synovial cells in articulationes sacroiliaca. The serum contents of TNF-α and DKK-1 were determined by ELISA,and the mRNA expressions of BMP-2,Cbfα1 and Smad1 in synovial tissue were detected by real-time fluorescent quantitative PCR. RESULTS:Compared with normal group,serum content of TNF-α in model group was increased significantly,while serum content of DKK-1 was decreased significantly;mRNA expressions of BMP-2,Cbfα1 and Smad1 in synovial tissue were increased significantly,with statistical significance (P<0.05 or P<0.01). Under the electron microscope,the synoviocytes of mice in model group were proliferated and arranged in disorder,and the secretory activity of active organelles were hyperactivity and the gap among cells became widened. Compared with model group, serum content of TNF-α in sulfasalazine group and paeonol group were decreased significantly;serum content of DKK-1 was increased significantly and mRNA expressions of BMP-2,Cbfα1 and Smad1 in synovial tissue were decreased significantly in paeonol group,with statistical significance (P<0.05 or P<0.01). Electron microscopy showed that mitochondria,lysosome and rough endoplasmic reticulum structure of synovial cell were improved significantly in paeonol group. CONCLUSIONS:The mechanism prevention and treatment of paeonol on AS may be associated with reducing serum content of TNF-α,increasing serum content of DKK-1,down-regulating mRNA expressions of BMP-2,Cbfα1 and Smad1 in synovial cells,inhibiting Wnt and BMP/Smad ossification related signal transduction pathway and reversing osteogenic differentiation of synovial cells.

Objective: To investigate the change in expression of anti-senescence marker protein calmodulin (RGN) in liver tissues of rats with immune hepatic fibrosis, and to observe the effect of compound glutathione inosine injection (CGII) on it. Methods: Rat liver fibrosis model was induced by intraperitoneal injection of porcine serum, and CGII intervention was administered at the appropriate time. Rat liver tissues were stained with HE and Masson. RGN and protein expression at mRNA in liver tissues was detected by fluorescence quantitative PCR and immunohistochemistry. One-way Anova was used for measurement data. LDS test was used for two-way comparison, and pathological semi-quantitative results were analyzed by rank-sum test. Results: The relative expression of RGN mRNA and protein in liver tissue of fibrotic rats was 82.23 ± 15.21 and 12.52 ± 3.23, respectively, which were significantly lower than that of normal rats 176.39 ± 11.35 and 59.23 ± 9.13 (P < 0.01). The degree of liver fibrosis in fibrotic rats after CGII intervention was significantly lower than fibrotic rats. The relative expression of RGN mRNA and protein in the intervention group was 168.78 ± 21.31 and 46.42 ± 4.71, respectively, which were significantly higher than fibrosis and spontaneous recovery group. The difference was statistically significant (P < 0.01). The relative expression of RGN mRNA and protein in the spontaneous recovery group was 86.23 ± 17.16 and 14.34 ± 5.16, which was higher than model group. The difference was not statistically significant (P > 0.05). Conclusion The expression of RGN in liver tissue of rats with hepatic fibrosis induced by porcine serum is decreased, while the expression of RGN increases with the decrease of fibrosis after CGII intervention, suggesting that the protein may play an important role in the development of liver fibrosis.

Objective: To investigate the effect and mechanism of XTP4 gene in apoptotic hepatoblastoma HepG2 cell line. Methods: HepG2 cells were transiently transfected with small interfering RNA of XTP4 genes, plasmid pcDNA3.1/myc-His(-) A-XTP4, and hepatitis B virus X protein transactivated x gene 4 (HBX protein trans-activate gene4, XTP4) and their respective negative controls. After 48h, the overexpression and interference expression condition of XTP4 in HepG2 cells were detected by Western blot. HepG2 cells apoptosis was detected by flow cytometry. The expression levels of apoptosis-related proteins P53, Bcl-2, Bax and Caspase-3 in HepG2 cells were detected by Western blot, and Bcl-2/Bax ratio was calculated. The chemiluminescence assay was used to detect activity of caspase-3 in HepG2 cells. The measured data were presented as ( ± s), and independent sample t-test was used for comparison between the two groups. Results: HepG2 cells had successfully achieved the overexpression and interference expression of XTP4 protein. Compared with the control group, the overexpression of XTP4 in HepG2 cells had significantly decreased the number of apoptotic cells (P < 0.05), and increased Bcl-2/Bax (P < 0.05) ratio, but decreased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was decreased (P < 0.05). However, interference with XTP4 expression in HepG2 cells had significantly increased the number of apoptotic cells (P < 0.05) and decreased Bcl-2/Bax (P < 0.05) ratio, but increased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was increased (P<0.05). Conclusion In HepG2 apoptosis XTP4 has inhibitory effect, and its effect on inhibiting HepG2 apoptosis may be achieved by regulating the Bcl-2/Bax ratio, and the P53 protein may be involved.

Objective To clone the murine Na+-K+-2Cl-cotransporter (Nkcc2) gene promoter and analyze 20-HETE regulation of the murine Nkcc2 gene transcriptional activity. Methods A fragment of the murine Nkcc2 gene promoter was analyzed using bioinformatics software AliBaba and TRANSFAC TESS. The murine Nkcc2 gene promoter fragment (-1 462 bp-+40 bp) was amplified by PCR using murine genomic DNA as a template and then cloned into a pGL3-Basic vector to generate a luciferase reporter construct. The recombinant reporter construct was transiently transfected into HEK293 T cells using Lipofectamine 2000 for 24 h. The transfected HEK293 T cells were treated with 20-HETE for 2 h followed by measurement of the luciferase activity using the Dual-Luciferase Reporter Assay system. Results A luciferase reporter construct containing the murine Nkcc2 gene promoter was successfully generated. The results showed that 20-HETE significantly reduced the transcriptional activity of the construct. Conclusion 20-HETE may reduce the expression of the murine Nkcc2 gene through transcriptional regulation.

Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found that high glucose(HG)levels in the cerebrospinal fluid(CSF)of diabetic rats might enhance the effect of a subthreshold dose of the neurotoxin 6-hydroxydopamine(6-OHDA)on the development of motor disorders,and the damage to the nigrostriatal dopaminergic neuronal pathway.In vitro,HG promoted the 6-OHDA-induced apoptosis in PC12 cells differentiated to neurons with nerve growth factor(NGF)(NGF-PC12).Metabolomics showed that HG promoted hyperglycolysis in neurons and impaired tricarboxylic acid cycle(TCA cycle)activity,which was closely related to abnormal mito-chondrial fusion,thus resulting in mitochondrial loss.Interestingly,HG-induced upregulation of pyruvate kinase M2(PKM2)combined with 6-OHDA exposure not only mediated glycolysis but also promoted abnormal mitochondrial fusion by upregulating the expression of MFN2 in NGF-PC12 cells.In addition,we found that PKM2 knockdown rescued the abnormal mitochondrial fusion and cell apoptosis induced by HG+6-OHDA.Furthermore,we found that shikonin(SK),an inhibitor of PKM2,restored the mito-chondrial number,promoted TCA cycle activity,reversed hyperglycolysis,enhanced the tolerance of cultured neurons to 6-OHDA,and reduced the risk of PD in diabetic rats.Overall,our results indicate that diabetes promotes hyperglycolysis and abnormal mitochondrial fusion in neurons through the upre-gulation of PKM2,leading to an increase in the vulnerability of dopaminergic neurons to 6-OHDA.Thus,the inhibition of PKM2 and restoration of mitochondrial metabolic homeostasis/pathways may prevent the occurrence and development of diabetic PD.

Objective:To explore the evaluation value of serum levels of positive pentameric protein 3 (PTX3) and creatine kinase isoenzyme MB (CK-MB) on volume load in patients with chronic decompensated heart failure (CDHF).Methods:A total of 300 CDHF patients who visited the Xingtai Central Hospital from July 2019 to July 2022 were selected and divided into a capacity overload group ( n=182) and a non capacity overload group ( n=118) based on their capacity balance level. Two clinical data sets were compared and analyzed. The receiver operating characteristic (ROC) curve was used to analyze the evaluation value of serum PTX3 and CK-MB levels on the volume load of CDHF patients. The clinical disease characteristics of the two groups of patients were analyzed using univariate analysis, and the influencing factors of volume load of CDHF patients were analyzed using logistic regression. A column chart model was constructed and validated. Results:The body mass index (BMI), waist circumference, systolic blood pressure, diastolic blood pressure, fasting blood glucose (FBG), glycosylated hemoglobin (HbA 1c), C-reactive protein (CRP), uric acid (UA), homeostasis model assessment of insulin resistance (HOMA-IR) of patients in the capacity overload group were higher than those in the non-capacity overload group, and the differences were statistically significant (all P<0.05). The PTX3, CK-MB, pulmonary capillary wedge pressure (PCWP), and CVP levels of patients in the capacity overload group were higher than those in the non-capacity overload group, while albumin, hemoglobin, and hematocrit were lower than those in the non-capacity overload group, and the differences were statistically significant (all P<0.05). The ROC curve showed that the area under the curve (AUC) of PTX3 and CK-MB for predicting capacity overload in CDHF patients are 0.795 and 0.718, with sensitivity of 86.2% and 83.7%, specificity of 65.4% and 68.6%, respectively, indicating high predictive accuracy; The AUC of the two joint predictions is 0.817, the sensitivity was 92.5%, and the specificity was 70.6%. The prediction accuracy was higher than PTX3 ( Z=3.812, P<0.05) and CK-MB ( Z=3.365, P<0.05). PTX3, CK-MB, albumin, hemoglobin, hematocrit, PCWP, and central venous pressure (CVP) were all influencing factors of volume load status in CDHF patients (all P<0.05). The column chart risk prediction model established based on these factors had high accuracy and strong applicability in clinical treatment. Conclusions:Serum PTX3 and CK-MB levels are influencing factors for volume overload in CDHF patients. A column chart model constructed in combination with indicators such as albumin, hemoglobin, hematocrit, PCWP, and CVP has high predictive value for the volume overload status of CDHF.