Objective To construct Neuropilins-2 eukaryotic expression vector for RNA interference.Methods Recombinant targeting on gene NRP2 was designed and established with plasmid pGenSil-1 based on NRP2 cDNA equences of Genomes.Two pairs of oligonucleotides were synthesized according to the Tuschl and inserted into plasmid pGenSil-l to generate siRNA eukaryotic expression vector,DH5? strains were transformed,plasmid were extracted,and recombinant vectors were identified by the restriction map and the sequence analysis.The recombinant plasmid(pGenSil-NRP2) was transfected into the cultured LOVO cells.At 48 h after transfection,the whole cell protein was extracted,and the protein level was detected by Western blotting with mouse-anti-human NRP2 monoclonal antibody.Results Recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis.pGenSil-NRP2 expression vector into LOVO cells down-regulated the protein level of NRP2 at 48 h after transfection.The recombinant eukaryotic expression vector were constructed successfully.Conclusion siRNA recombinant can be constructed successfully by RNAi technique for inhibiting NRP2 expression.

Objective To investigate the current prevailing status of endemic fluorosis in the Yellow River basin of Shandong Province and to provide the scientific evidence for making strategies in prevention and control.Methods Nine counties were chosen to carry out the epidemiological investigation.The content of fluoride in drinking water was determined by F-ion selective electrode and dental fluorosis of children aged 8～12 years old was diagnosed by Deans method.Results Water fluoride content was determined in 1761 fluorosis villages,among which 606 villages had water fluoride content≤1.00 mg/L,accounting for 34.41%(606/1761);1155 villages had water fluoride content＞1.00 mg/L,which accounted for 65.59%(1155/1761).The highest water fluoride content was 11.33 mg/L.Water fluoride content of 618 water-improving and defluoridation projects had been determined,among which 449 projects had water fluoride content≤1.00 mg/L and accounted for 72.65%(449/618),169 projects had water fluoride content＞1.00 mg/L and accounted for 27.35%(169/618),the highest water fluoride content was 5.85 mg/L.The total rate of dental fluorosis of children aged 8～12 years old was 45.03%(25 579/56 804) and the index of dental fluorosis was 0.80. Conclusions In the Yellow River basin in Shandong Province,up to 50.00%in the villages the water fluoride content exceeds the county standard(≤1.00 mg/L).The prevalence of endemic fluorosis in the basin hasn't been effectively controlled.So the counterrneasures for endemic fluorosis should be carried out as soon as possible.

Ultrasound therapy is a new type of therapy technology, there are hyperthermia and high intensity focused ultrasound surgery (HIFUS). Compared with a single frequency system, mutiple frequency system has an additional function to combine power patterns of different frequencies. This function increases the availability of power patterns to treat tumors of various shapes and depths. Therefore, we have in this article proposed a system with the ability to drive ultrasonic phased arrays of multiple resonant frequencies for ultrasound hyperthermia and HIFUS. The results show that this system is able to (1) drive multi-element applicators or phased arrays of a single resonant frequency through the multichannel power amplifiers, (2) concurrently drive transducer with different resonant frequencies, (3) adjust the relative phase and output power of each channel for the scanning ultrasonic focus, and (4) operate each channel with good output stability.
Equipment Design ; Humans ; Neoplasms ; therapy ; Transducers ; Ultrasonic Therapy ; instrumentation

Objective To investigate selcnium(Se) levels of environment and human body in Gaomi City and Zichuan District of Shandong. Methods Lijiaying Township in Gaomi City of Weifang City, Zhaili Township and Longquan Township in Zichuan District of Zibo City were selected. Two farming soil samples at different spot, local wheat and corn, residents nail samples from 3 to 4 families were collected in each natural village in the investigated towns. The contents of Se were detected by 2,3-diamino naphthalene fluorescence method. Results Se level of the soil, wheat, corn, and nails in Lijiaying [(0.054 ± 0.019), (0.022 ± 0.009), (0.018 ± 0.007), (0.365 ± 0.108)mg/kg] was significantly lower than that in Zhaili [(0.425 ± 0.080), (0.130 ± 0.043), (0.098 ± 0.026), (0.751 ± 0.134)mg/kg] and Longquan[(0.487 ± 0.153), (0.112 ± 0.030), (0.097 ± 0.029), (0.735 ± 0.145)mg/kg;P < 0.01]. In Lijiaying, Se was deficient in soil, wheat, corn(< 0.200, < 0.025 mg/kg), above Se deficiency diagnosis and below Se-adequate level in the nail, while in Zhaili and Longquan, the Se level in the soil (0.425, 0.487 mg/kg), wheat(0.130, 0.112 mg/kg), corn (0.098, 0.097 mg/kg), nails (0.751, 0.735 mg/kg) was adequate (≥0.400 mg/kg). Conclusions The external environment is Se-deficient in Lijiaying, Se-adequate in Longquan and Zhaili. The selenium level in human body is consistent with the external environment.

Objective: To investigate the clinicopathologic characteristics and BRAF V600E mutation of brain tumors associated with epilepsy. Methods: Totally 250 patients with brain tumors associated with epilepsy were included from March 2008 to August 2017 retrospectively at Sanbo Brain Hospital, Capital Medical University.The clinical manifestations, histological features and BRAF V600E mutation results were collected and analyzed. Results: There were 132 males and 118 females, and the male to female ratio was 1.1∶1.0. The age of patients ranged from 2 to 67 years(mean 22 years). The tumors had obvious local space occupying effect on MRI. The temporal lobe was the most common site (44.4%, 111/250). There were 58.4% (146/250) of ganglioglioma (GG), 24.0% (60/250) of dysembryoplastic neuroepithelial tumor (DNT), 12.8% (32/250) of pleomorphic xanthoastrocytoma(PXA), 4.0% (10/250) of angiocentric glioma (AG) and 0.8% (2/250) of papillary glioneuronal tumor (PGNT). Mixed GG, PXA and DNT morphological structures were found in 9 of patients. Among 250 cases, 35 cases were accompanied by focal cortical dysplasia(FCD). BRAF V600E was seen in 43 of 74 (58.1%) GG and 13 of 28 (46.4%) PXA. The most common pathologic grade of GG, DNT, AG and PGNT was WHO I. Some of the tumor cells from GG (34 cases) showed higher proliferative activity (WHO Ⅱ/Ⅲ). Most cases of PXA were WHOⅡand high proliferative activity was seen in nine cases. Conclusions The association of low-grade glioneuronal tumors with intractable epilepsy was well-recognized. The most common low-grade glioneuronal tumors were GG.GG may occur in any part of the central nervous system, with a predilection for temporal lobe. Each type of low-grade glioneuronal tumors has its own unique histological morphology, but some may show complex features with 2 or 3 mixed components. The occurrence of BRAF V600E mutations in GG is common, and their detection may be valuable for the diagnosis and treatment in GG.

Objective:To analyse the characteristics of micronuclus and chromosomal aberrations and the lens opacity in different working years, ages and work type of radiation workers with lens opacity.Methods:In January 2022, we selected 400 radiation workers with lens opacity who had undergone occupational health examination in Hangzhou Hospital for the Prevention and Treatment of Occupational Disease from March 2014 to December 2021 as research objects. To analyze the differences of micronucleus, chromosome aberration and the lens opacification rates of different length of service, type of work, age radiologic workers.Results:The results showed that radiation the micronucleus abnormality (0.59‰) and chromosome aberration (2.67%) in peripheral blood in workers who served more than 20 years were significantly higher than that of micronucleus abnormality (0.18‰) and chromosome aberration (0.31%) in workers served less than 20 years, the differences were statistically significant ( P<0.05). The micronucleus abnormality (0.32‰) and chromosome aberration (0.57%) in the interventionist were higher than oral radiation workers (0.12‰ and 0.19%) and diagnostic radiologist (0.05‰ and 0.12%), the differences were statistically signcficant (χ 2=23.98, 6.72, P<0.05) . Conclusion:The rates of micronucleus and chromosome aberration of the interventionaist are higher than oral radiation workers and the traditional radiologist. Workers engaged in radiation should improve personal protection and undertake regular occupational health examinations.

The cross reactivity of mimotopes of hepatitis C virus (HCV) hypervariable region 1 (HVR1) was investigated to obtain epitopes that have high cross reactivity. Five expression vectors encoding B cell mimotopes fused with Trx were constructed, and the mimotope proteins were purified. The cross reactivity of mimotope proteins with HCV positive sera was determined by ELISA. HCV pseudotype particles (HCVpp) were generated and applied to evaluate neutralization effects of the sera of BALB/c mice immuned with the mimotope proteins on infection of Huh7. 5 cells. Our data showed that the mimotope proteins (P1, P2, P5, P6, P8) could react to the HCV positive sera. The HCVpp infection inhibition of the sera of BALB/c mice immuned with P6 or P8 was detectable. These results suggest that the mimotopes may be valuable in the studies of anti-HCV infection and development of HCV vaccines.
Amino Acid Sequence ; Animals ; Complementarity Determining Regions ; immunology ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; Hepacivirus ; immunology ; Humans ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data

OBJECTIVETo investigate the effect of interferon regulatory factors (IRFs) on neointimal formation after vascular injury in the mouse, and its possible mechanism.

METHODSVascular injury was induced by polyethylene cuff placement around the left femoral artery of IRF-1-deficient mice and C57BL/6J mice. The mRNA expressions of IRF-1, IRF-2, angiotensin II type 2 (AT2) receptor, interleukin-1 beta converting enzyme (ICE), inducible nitric oxide synthase (iNOS) were detected by RT-PCR and immunohistochemical staining.

RESULTSNeointimal formation after vascular injury was significantly greater in IRF-1-deficient mice than that in C57BL/6J mice (P<0.05). In contrast, TUNEL-positive nuclei to total nuclei in the neointima and media in vascular smooth muscle cell (VSMC) in the injured artery significantly attenuated in IRF-1-deficient mice compared to C57BL/6J mice (P<0.05). The expressions of AT2 receptor as well as pro-apoptotic genes such as ICE and iNOS in C57BL/6J mice were up-regulated in response to vascular injury, but this upregulation was attenuated in IRF-1-deficient mice.

CONCLUSIONSOur results suggest that IRF-1 induces VSMC apoptosis and inhibits neointimal formation after vascular injury at least partly due to the upregulation of AT2 receptor, ICE and iNOS expressions. These results indicate that IRF-1 exerts an inhibitory effect on neointimal formation through the induction of apoptosis in VSMCs.

Animals ; Apoptosis ; physiology ; Caspase 1 ; genetics ; metabolism ; Femoral Artery ; anatomy & histology ; pathology ; Interferon Regulatory Factor-1 ; genetics ; metabolism ; Interferon Regulatory Factor-2 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle, Smooth, Vascular ; cytology ; metabolism ; pathology ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; genetics ; metabolism ; Receptor, Angiotensin, Type 2 ; genetics ; metabolism ; Tunica Intima ; pathology ; physiology

BACKGROUNDSchwannoma is the tumor arising mainly from the cranial and spinal nerves. Bilateral vestibular schwannoma is the hallmark of neurofibromatosis type 2 (NF2). The NF2 gene has been cloned with comprehensive analysis of its mutations in schwannoma. However, most studies focused on vestibular schwannoma. There are differences in proliferation of tumor cell and ultrastructure between vestibular and spinal schwannomas. It is unknown whether genetic alterations in vestibular schwannoma are different from those in non-vestibular schwannoma. We analyzed the loss of heterozygosity (LOH) on chromosome 22 in patients with sporadic schwannoma including vestibular and spinal schwannomas and correlated this genetic alteration with tumor proliferation.

METHODSIn 54 unrelated patients without clinical NF1 or NF2, 36 patients had sporadic vestibular schwannoma, and 18 dorsal spinal root schwannoma. Four highly polymorphic linkage to NF2 gene microsatellite DNA markers (D22S264, D22S268, D22S280, CRYB2) were used to analyze LOH. The proliferative index was evaluated by Ki-67 and proliferative cell nuclear antigen (PCNA) immunostaining. Student's t test was used to analyze the difference of the proliferative index between schwannoma with LOH and that without LOH. The difference of the frequency of LOH in vestibular and spinal schwannomas was investigated by the chi-square test.

RESULTSTwenty-three schwannomas (42.6%, 23/54) showed allele loss. The frequency of LOH in vestibular schwannoma was significantly higher than that in spinal schwannoma (chi2 = 5.14, P < 0.05). The proliferative index of schwannoma with LOH was significantly higher than that without LOH (tki-67 = 2.97, P = 0.0045; tPCNA = 2.93, P = 0.0051).

CONCLUSIONSLOH on chromosome 22 is a frequent event in the tumorigenesis of sporadic schwannoma. And, there is a correlation between LOH on chromosome 22 and proliferative activity in schwannoma. The frequency of LOH in vestibular schwannoma is significantly different from that in spinal schwannoma.

Adult ; Aged ; Cell Proliferation ; Chromosomes, Human, Pair 22 ; Female ; Genes, Neurofibromatosis 2 ; Humans ; Loss of Heterozygosity ; Male ; Middle Aged ; Neurilemmoma ; genetics ; pathology ; Neuroma, Acoustic ; genetics ; Spinal Cord Neoplasms ; genetics ; Spinal Nerve Roots

Objective To explore the correlation between environmental selenium(Se) levels and cognitive ability among rural elderly population, and the effect of Se on cognitive skill of the elderly. Methods Two study sites(Zichuan district and Gaomi city of Shandong) with different environmental Se levels were selected according to rural elderly people ≥ 65 years were extracted by stratified random sampling method in each site. A retrospective survey was carried out using dietary intake questionnaire for the elderly for the past 1 year, and their daily total Se intake was calculated. Questionnaire was also used to obtain cognitive skill information in the elderly people. The Community Screening Instrument for Dementia, the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) Word List Learning Test, the Indiana University (IU) Story Recall Test, the Animal Fluency Test, and the IU Token Test were applied to assess the cognitive ability. Se level in these samples was analyzed. Correlation between the Se levels of environment and those of human body were analyzed statistically. Results There were 0.163), (0.405 ± 0.086)]mg/kg and nail Se[(147.44 ± 17.42), (117.38 ± 22.48)μg/L]between the two groups (U = 31.59, 25.00, 23.67, all P ＜ 0.01 ). There were positive correlation among the nail Se, environmental Se and Se in all subjects(r = 0.51,0.46, 0.60, all P ＜ 0.01 ). The differences of the CSID total score, the CERAD Word List Learning Test, the CERAD Word List Recall Test and the IU Story Recall Test between the two sites were statistically significant(F = 2.56, 9.18, 7.48, 4.42, all P ＜ 0.05), excluding the Indiana University Story Recall Test.After eliminating possible confounding factors, the Se levels and the CSID total score, the IU Story Recall Test, the Animal Fluency Test and the IU Story Token Test had a significantly positive correlation (r = 0.076, 0.138, 0.042,0.107, P ＜ 0.05 or ＜ 0.01 ), excluding the CERAD Word List Learning Test. Conclusions This study supports the hypothesis that a life long low Se level is associated with lower cognitive ability. The cognitive abilities in the elderly population lived in areas with high environmental selenium levels are significantly higher than that of the elderly lived in areas with low environmental selenium levels.