Changes of cardiac metabolite profile in offspring rats under prenatal stress / 西安交通大学学报(医学版)

【Objective】 To investigate the effect of prenatal stress (PS) on the profile changes of cardiac metabolites in offspring rats and to analyze the potential role of key differentiators in key differential signaling pathways. 【Methods】 UHPLC-Q-TOF/MS analysis was used to detect the changes of metabolite profile in the heart tissues of offspring rats in control group and PS group. KEGG pathway annotation and analysis were used to screen out metabolic differences in key signaling pathways and quantify their expression levels so as to predict the potential function of these key molecules in the effect of PS on the heart tissues of offspring rats. 【Results】 Compared with the control group rats, the signaling pathways in the PS offspring rats’ heart tissue that changed significantly included biosynthesis of amino acids, purine metabolism, pyrimidine metabolism, alanine, aspartate and glutamate metabolism, cAMP signaling pathways, arginine biosynthesis, GABAergic synapses, glutamate synapse, nicotine addiction, and regulation of actin cytoskeleton. Among them, the levels of L-glutamine, pseuduracil, uric acid, xanthine, 2’-deoxyadenosine 5’-monophosphate, cytosine 3’-monophosphate, and cytosine 5’-monophosphate were upregulated, while the level of argininosuccinic acid was downregulated, which enriched in purine metabolism, pyrimidine metabolism, and arginine biosynthesis pathway. 【Conclusion】 PS leads to abnormal changes of L-glutamine, pseuduracil, uric acid, and xanthine in the heart tissue of offspring rats, and PS may be a high risk factor for cardiovascular diseases in offspring rats.

A novel modified scleral tunnel prevents tube exposure in Ahmed valve implantation / 眼科新进展

Objective To evaluate the effectiveness and safety of modified scleral tunnel for the prevention of Ahmed valve tube exposure in Ahmed valve implantation for refractory glaucoma.Methods A retrospective study was conducted in 72 patients with Ahmed glaucoma valve implantation for refractory glaucoma.All the patients were divided into unmodified and modified scleral tunnel group.In the unmodified scleral tunnel group,38 patients (38 eyes) underwent traditional scleral tunnel in Ahmed valve implantation,while in the modified scleral tunnel group,34 patients (34 eyes) received modified scleral tunnel in the procedures.The changes in intraocular pressure and visual acuity after operation were observed,and the incidence of drainage tube exposure and other complications were compared between the two methods after Ahmed valve implantation.The patients were followed up for 6 to 36 months with an average of 18 months.Results The intraocular pressure was (16.30 ±5.73) mmHg (1kPa =7.5 mmHg) in the unmodified scleral tunnel group and (15.80 ± 6.12)mmHg in the modified scleral tunnel group,with no significant difference between the two groups (P ＞ 0.05),but there was significant difference with the preoperative data (48.4 ± 5.79) mmHg (P ＜ 0.01).During the follow-up,conjunctival tube exposure was seen in 3 eyes (7.9％) in the unmodified scleral tunnel group,whereas there was no tube exposure in the modified scleral tunnel group.Conclusion The modified scleral tunnel is capable to prevent conjunctival tube exposure in patients with refractory glaucoma,which is more efficient and safe than traditional scleral tunnel in Ahmed glaucoma valve implantation.

Expression purification and clinical application of high-risk HPV16 E4 gene / 中南大学学报(医学版)

OBJECTIVE@#To determine the relationship between serum antibody against HPV16 E4 and cervical cancer, and to construct HPV 16 E4 protein expression vector as an antigen to detect its corresponding serum antibody among different populations.@*METHODS@#HPV16 E4 early gene was ligated into pRSET-A expression vector. The constructed plasmids were transformed into BL21 (DE3)cells, and induced to express HPV 16 E4 protein by isopropylthio-beta-D-galactoside (IPTG). The expressed E4 inclusions were denatured, purified through Ni-column, and renatured. After the activity was revealed, antibodies against HPV 16 E4 in the sera from healthy women and patients with chronic cervicitis and cervical cancer were respectively determined by enzyme-linked immunosorbent assay (ELISA) using the fusion protein as the antigen.@*RESULTS@#HPV 16 E4 fusion protein of Mr 15*10(3) was expressed by pRSET-16E4 after IPTG induction. The fusion protein accounted for 30% of the total bacterial proteins and expressed as inclusive body. After purification with Ni-NTA agarose resin, the recombinant protein revealed purity of 95%, and activity of the renatured protein was identified by ELISA. The serum antibody-positive rate of HPV 16 E4 was 10.00%, 39.13% and 28.13%, respectively in 80 healthy women, 46 chronic cervicitis patients, and 32 cervical cancer patients. The antibody-positive rate in cervical cancer patients and chronic cervicitis patients were significantly higher than that in healthy women (P<0.01), while the difference between the antibody-positive rate in cervical cancer patients and chronic cervicitis patients was not significant.@*CONCLUSION@#HPV 16 E4 protein expressed from pRSET-A/BL21 can be used in serological studies on cervical cancer-related HPV infection. Serum antibody against HPV16 E4 is present in a significantly higher percentage in cervical cancer and chronic cervicitis patients than in healthy women.

Detection of HPV 58 and cloning of its E7 gene in cervical cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect HPV 58, a common type of human papillomavirus (HPV), clone and express its E7 gene from biopsy specimens of cervical cancer.</p><p><b>METHODS</b>HPV 58 from 58 biopsy tissues of cervical cancer was detected by GP5+/GP6+ PCR followed by template-directed dye-terminator incorporation assay with fluorescence polarization detection (TDI-FP). HPV 58 E7 gene was amplified from one HPV 58-positive sample, and then cloned into pGEM-T Easy vector. The recombinant plasmid, HPV58E7-pGEM-T was confirmed by sequencing. Subsequently, E7 gene was cloned into prokaryotic expression vector pRSET-A. The constructed pRSET-58E7 plasmids were transfected into BL21(DE3) cells, and induced to express 58 E7 protein by IPTG.</p><p><b>RESULTS</b>Among the 58 biopsy tissues of cervical cancer, 10 were HPV 58-positive, accounting for 19.2% of 52 HPV-positive cases. HPV 58 E7 gene was amplified from one HPV 58-positive sample. The constructed plasmids were identified containing HPV58 E7 gene by restriction enzyme analysis and sequencing. SDS-PAGE analysis showed that HPV58 E7 His6 fusion protein of M(r) 16 x 10(3) was expressed by pRSET-58E7 after induction by IPTG. The fusion protein accounted for 30% of total bacterial proteins.</p><p><b>CONCLUSION</b>HPV 58 is not uncommon in Chinese women with cervical cancer in Shaanxi province. Constructed HPV58 E7 recombinant plasmids can be effectively expressed in E.coli, which may provide a tool in diagnosis and vaccine design for HPV of HPV58-associated tumors.</p>

A high throughout assay for human papillomavirus genotypes with fluorescence polarization / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To develop a simple, cheap, quick, accurate and practical method for a high throughout genotypes assay of human papillomavirus (HPV) DNA.</p><p><b>METHODS</b>Crude DNA was extracted by a simplified proteinase K digesting method. HPV common conservative primers: GP5+/6+ system was used to amplify HPV DNA in 127 samples of condylomata acuminatum (CA) and cervical scrapes by PCR, then the PCR product was assayed using a template directing terminator incorporation (TDI) and genotypes were detected with fluorescence polarization (FP). Major HPVs type-specific probes (HPV6, 11, 16, 18, 31, 33, 35 and 58) designed by us were hybridized with the specific PCR products and a special fluorescent ddNTP terminator was directly added to the end of the probe under direction of specific PCR products. The results were measured with FP and compared with the results of the DNA sequence.</p><p><b>RESULTS</b>Compared with the results of DNA sequencing, the results detected with fluorescence polarization were all correct. The proposed method could detect more than one type of HPV infection, but DNA sequencing method could not. The positive rate of HPV was 100% in 78 CA biopsies. Among them, there were 14 HPV double infections [HPV6B and 11 (9 cases), HPV11 and 16 (4), HPV11 and 18 (1)], 5 HPV triple infections [HPV6B, 11 and 16 (4), HPV11, 16 and 18 (1)], and one HPV quadruple infection (HPV6B, 11, 16 and 18). The positive rate of HPV was 77% in the 49 cervical scrapes. Six HPV double infections [HPV6B and 11 (2), HPV11 and 16 (1), HPV6B and 16 (1), HPV16 and 18 (1), HPV18 and 58 (1)], 3 HPV triple infections [HPV6B, 11 and 16 (2), HPV11, 16 and 18 (1)] and one HPV quadruple infection (HPV6B, 11, 16 and 18) were detected in cervical cancer scrapes.</p><p><b>CONCLUSIONS</b>The proposed method allowed a high throughout, special, simple, rapid, automatic and economical detection of HPV-DNA genotyping without a use of labeling probes. It can detect multiple HPV genotype infection and will be and useful tool in HPV genotype screening.</p>