ACEI,ACEII and Xyr1 are transcriptional factors that regulate cellulase gene expression in Trichoderma koningii.In vitro experiments have shown that ACEI and Xyr1 bind to the cbh1 promoter fragment(-304 to-18) and regulate the gene transcription.However,whether ACEII binds to this 287bp fragment is still unclear.To further elucidate the regulatory mechanism of ACEII for cellulase,DNA-binding domains of ACEII from T.Koningii were expressed in E.coli.It could not show binding to the cbh1 promoter fragment(-304 to-18) by electrophoresis mobility shift assays,suggesting that it is Xyr1 but not ACEII binds playing an essential role during induction of cbh1 gene transcription.

Objective　To investigate the role of bacterial DNA in systemic inflammatory response syndrome (SIRS). Methods　A total of 100 mice of Kunming species were divided into ten groups: E.coli DNA (30, 20, 10, 5 and 1 mg/kg ), 30 mg/kg of CT DNA, 60Co DNA, DNased DNA, organic residue of DNA extraction and sterile water control. The last two were pre-treated with D-galactoamine (600 mg/kg intra peritoneally). Animals were administratively injected via tail vein. General physical condition and the death rate of mice were observed within 48 h. Results　①Obvious lethal effect of double strand E.coli DNA on mice were observed with a dose-effect correlation, LD50=11.51 mg/kg. ②NO difference in death rate was found in the group of 30 mg/kg E.coli DNA with or without 60Co irradiation (10/10 and 8/10,P>0.05). ③No rats died in the group of DNased DNA, organic residue of DNA extraction and calf thymic DNA (0/10). Conclusion　Bacterial DNA may play an important role in the development of SIRS.

Objective To investigate the relationship between angiotensin Ⅱ and pancreatic islet β cell secretion function under different glucose tolerance statuses. Method Forty-two patients with newly diagnosed type 2diabetes mellitus ( DM group), 38 subjects with impaired fasting glucose/impaired glucose tolerance ( IFG/IGTgroup) ,and 40 normal control subjects (NGT group) underwent intravenous glucose tolerance test. Fasting plasma angiotensin Ⅱ ( Ang Ⅱ ) and adiponectin were assayed by ELISA. Acute insulin response from 3 to 10 min( AIR3-10 ),the area under the curve( AUCⅠ ) and the peak concentration of the first-phase ( 0-10 min) insulin secretion, the area under the curve of the second-phase( 10-120 min) insulin secretion( AUCⅡ), homeostasis model assessment for β cell function index(HOMA-β) and homeostasis model assessment for insulin resistance index(HOMA-IR) were calculated to explore the relationship with Ang Ⅱ. Result ( 1 ) The levels of Ang Ⅱ in DM group and IFG/IGT group were significantly higher than that in NGT group( P＜0.05 ). The AIR3-10, AUCⅠ and peak concentration, AUCⅡ ,adiponectin in DM group and IFG/IGT group were significantly lower than those in the NGT group ( P＜0. 05), and these results were more significantly reduced in DM group compared with those in IFG/IGT group. (2) Ang Ⅱ was negatively correlated with AIR3-10, AUCⅠ and the peak concentration, AUCⅡ, adiponectin, HOMA-β ( P＜0. 01 ), and positively correlated with fasting blood glucose,2 h blood glucose after glucose loading, fasting insulin, HOMA-IR (P＜0. 05 ). (3)Multiple stepwise regression analysis showed that Ang Ⅱ was independently associated with AUCⅠ and AUCⅡ.Conclusion Ang Ⅱ was an independent factor that affected the insulin secretion function of pancreatic islet βcells. Ruling out the effect of blood pressure, body position, drugs, and other factors, high levels of Ang Ⅱ could predict the dysfunction of pancreatic islet β cell as well as insulin resistance in patients with type 2 diabetes.

The relationship between oxidative stress and the first-phase of pancreatic β cell insulin secretion in subjects with different statuses of glucose tolerance was explored. Fasting adiponectin, 8-hydroxydeoxyguanosin ( 8-OHdG), malondialdehyde ( MDA ), superoxide dismutase ( SOD ), insulin area under the curve ( AUC ) from 0 to 10 min, and AIR3-5 were measured. The levels of oxidative stress-related markers were elevated, the activities of superoxide dismutase,adiponectin, and first-phase of insulin secretion were reduced with the disease progression. MDA and 8-OHdG were negatively correlated with adiponectin, homeostasis model assessment β cell function index ( HOMA-β ),AUC ,and AIR3-5. SOD was positively correlated with adiponectin, HOMA-β, AUC, AIR3-5. The plasma 8-OHdG and SOD were independently associated with AIR3-5. Oxidative stress exerts a significant effect on the first phase of pancreatic β cell insulin secretion, which may contribute to the pathogenesis of type 2 diabetes.

Objective To investigate the relationship between adiponectin and the first-phase of pancreatic P-cell insulin secretion in subjects with different statuses of glucose tolerance. Methods Thirty-seven patients with newly diagnosed type 2 diabetes mellitus (DM) , 30 patients with abnormal glucose tolerance (IGR) , and 40 normal control subjects (NGT) underwent intravenous glucose tolerance test (IVGTT). Fasting adiponectin and proinsulin (PI) was assayed by EL1SA. Fasting free fatty acid ( FFA) was measured by colorimetry. Insulin area under the curve ( AUC ) , incremental AUC (iAUC) from 0 min to 10 min, AIR3-5, homeostasis model assessment for insnlin resistance (HOMA-IR) , and for β cell function ( HOMA-p) were calculated. The relationship between adiponectin and AUC, iAUC, AIR3-5, proinsulin, FFA, and HOMA-IR was explored. Results (1) The levels of AUC, iAUC, AIR3-5, and adiponectin in DM group and IGR group were significantly lower than those in NGT group (P<0.05), reduced in DM group than those in IGR group(P<0.05). (2) The levels of PI in DM group and IGR group were significantly higher than that in NGT (P<0.05). (3) Adiponectin was positively correlated with HOMA-p,AUC,iAUC,AIR3-5, and HDL-C,while negatively correlated with proinsulin, HOMA-IR, and LDL-C. (4) Proinsulin was positively correlated with HOMA-IR. (5 ) Multiple regression stepwise analysis showed that adiponectin was independently associated with AUC. Conclusions Adiponectin was an independent factor affecting the first phase of pancreatic p-cell insulin secretion. Low adiponectin level could predict the dysfunction of the first phase pancreatic p-cell secretion as well as insulin resistance in patients with type 2 diabetes.

ACEI and Xyr1 are two regulators that potentially involve in regulation of cellulases and xylanases formation in Trichoderma reesei, they compete for a binding site in the xyn1 (Xylanase1-encoding) gene promoter. To further investigate the mechanism for the transcriptional regulation of cellulases, DNA-binding domains of both ACEI and Xyr1 in T. Koningii were expressed from E. coli. They both showed bindings to the cbh1 promoter fragment (-304 bp to -18 bp) by electrophoresis mobility shift assays, suggesting ACEI and Xyr1 not only compete for binding to xyn1 promoter but also to cbh1 promoter.
Binding Sites ; Cellulase ; biosynthesis ; Endo-1,4-beta Xylanases ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Fungal ; Promoter Regions, Genetic ; Trans-Activators ; genetics ; metabolism ; Trichoderma ; enzymology ; genetics

Objective:To understand the molecular transmission characteristics of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome(AIDS) patients in the mountainous area of southwest Zhejiang Province(Lishui city).Methods:A total of 147 blood samples were collected from newly-diagnosed HIV-1 infected who received no antiviral therapy, and pol gene was amplified, followed by sequencing. MEGA6.0 software was used to construct phylogenetic tree and determine gene subtypes. HIVDB online was used to analyze drug resistance mutation, then the pairwise genetic distance(GD) was calculated and the opitimal threshold of GD was selected, finally the molecular transmission network was constructed by Cytoscape3.7.0 software. Chi-square or Fisher′s exact probability method was used for statistical analysis. Results:A total of 134 sequences were obtained successfully, and nine subtypes were detected. The dominant subtypes were CRF08_BC (34.33%, 46/134), CRF01_AE (29.85%, 40/134) and CRF07_BC (23.88%, 32/134). It also found that age, registered residence, education level and transmission route had significant differences in distribution of subtypes ( P<0.05). Nineteen drug resistance individuals were found, and the total drug resistance rate was 14.18% (19/134). The HIV-1 molecular transmission network was plotted based on 1.2% GD threshold. A total of 15 transmission clusters (cluster size ranging from 2 to 29) were found. The network access rate was 49.25% (66/134), mainly including male (75.76%, 50/66), heterosexual (81.82%, 54/66) and patientsrinfected with CRF08_ BC (50.00%, 33/66). A transmission cluster including two cases of female sex workers and seven cases of drug resistance was identified, in which the average age of the patients was 57.21 years old and the average degree value was 22.7, and the cases were mainly infected through heterosexual contact (96.55%, 28/29). The highest homology of the sequences in the cluster was in Yunnan. Conclusions:The HIV-1 subtypes were diverse in the mountainous area of southwest Zhejiang Province(Lishui city). Drug resistant transmission had reached a moderate epidemic level. There were molecular transmission clusters with the aggregation characteristics of elderly clients in specific regions. It was urgent to formulate and implement precise intervention strategies to curb the spread of HIV.

Objective:To explore the application value of chromosome karyotype analysis, chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in prenatal diagnosis of isolated corpus callosum abnormality (CCA) fetus.Methods:Fetuses diagnosed with isolated CCA by ultrasound and MRI and receiving invasive prenatal diagnosis in Guangzhou Women and Children′s Medical Center and Qingyuan People′s Hospital from January 2010 to April 2021 were selected. Karyotype analysis and/or CMA [or copy number variation sequencing (CNV-seq)] were performed on all fetal samples, and WES was performed on fetal samples and their parents whose karyotype analysis and/or CMA (or CNV-seq) results were not abnormal.Results:Among 65 fetuses with isolated CCA, 38 cases underwent karyotype analysis, and 3 cases were detected with abnormal karyotypes, with a detection rate of 8% (3/38). A total of 49 fetuses with isolated CCA underwent CMA (or CNV-seq) detection, and 6 cases of pathogenic CNV were detected, the detection rate was 12% (6/49). Among them, the karyotype analysis results were abnormal, and the detection rate of further CMA detection was 1/1. The karyotype results were normal, and the detection rate of further CMA (or CNV-seq) detection was 14% (3/21). The detection rate of CMA as the first-line detection technique was 7% (2/27). A total of 25 fetuses with isolated CCA with negative results of karyotyping and/or CMA were tested by WES, and 9 cases (36%, 9/25) were detected with pathogenic genes. The gradient genetic diagnosis of chromosomal karyotyping, CMA and WES resulted in a definite genetic diagnosis of 26% (17/65) of isolated CCA fetuses.Conclusions:Prenatal genetic diagnosis of isolated CCA fetuses is of great clinical significance. The detection rate of CMA is higher than that of traditional karyotyping. CMA detection could be used as a first-line detection technique for fetuses with isolated CCA. WES could increase the pathogenicity detection rate of fetuses with isolated CCA when karyotype analysis and/or CMA test results are negative.