Background N-ethyl-N-nitrosourea (ENU)-induced mouse mutagenesis is a powerful approach for the study of gene function and the generation of human disease models.Objective This study was to create the corneal morphologic change and map the mutant gene of a kind of corneal opacity in ENU mutagenesis in mouse.Methods ENU was intraperitoneally injected in forty C57BL/ 6J (B6) male mice aged 8-10 weeks old.The male mice were mated with the same strain female mice.Their progenies were screened for visible eye mutation,and the mutant mice were mated with the same strain mice to confirm the heredity of mutation phenotypes.Hematoxylin & eosin staining was used to examine the histopathological change of cornea in one mouse with ENU-induced corneal opacity.To map the mutant gene,[(B6×D2)F1 ×B6] N2 mutant mice were bred,and the genome of the N2 mice was scanned by microsatellite markers distributed equally on the mouse chromosome.The microsatellite linked to the mutant gene was determined by the log odds score.This experimental procedure was approved by Ethic Committee about Experimental Animal Care and Use of Yangzhou University.Results The founder mouse,which was the progeny of an ENU-treated B6 male mouse and an untreated B6 female mouse,had a corneal opacity phenotype.After mating the mutant with B6 mice,19 of 59 descendants appeared corneal opacity phenotype.Thickening of corneal stroma,neoangiogenesis,infiltration of inflammatory cells and proliferation of fibroblasts were exhibited in cloudy cornea in ENU-induced mutated mice under the optical microscope.After linkage analysis between microsatellite markers and the mutant gene,the mutant gene was linked to D2Mi307,which was located at 63.42 cM.Three cases of 26 N2 mice underwent recombination with the LOD 3.79.The mutant gene associated with the cornea phenotype was located on chromosome 2.Conclusions This study map the mutant gene associated with the cornea phenotype on chromosome 2.The strain might be used as a mouse model for heritable human corneal opacity.

BACKGROUND: The immediate implantation in the anterior maxillary region is in a high risk of aesthetic complications. OBJECTIVE: To assess the prognosis of the labial bone plate after anterior maxillary repair with immediate implant combined with immediate restoration. METHODS: Thirty-two patients with single failed tooth in the anterior maxillary region were subjected to implantation of ZIMMER implants immediately after minimally invasive extraction. Good primary stability was achieved and immediate restoration was carried out. Final restoration was finished after 6-12 months of osteosynthesis and gingival shaping. The loading situation of the labial bone plate was recorded at 6 months post operation. RESULTS AND CONCLUSION: Final restoration was finished with normal loading in all the patients. No bleeding and swelling of the gingiva was recorded. The horizontal absorption of the labial bone plate at the upper margin, 5 mm and 10 mm below the upper margin was (-2.12±0.05), (-1.54±0.04), and (-1.01±0.06) mm, respectively. Therefore, absorption of the labial bone plate with varying degrees exists after anterior maxillary repair with immediate implant combined with immediate restoration.

OBJECTIVETo investigate the relationship between vascular endothelial growth factor D (VEGF-D) and metastasis of lymphatic vessel in gastric carcinoma.

METHODSThe human VEGF-D cDNA was amplified from total RNA isolated from human normal gastric tissue, then it was inserted into T-A clone plasmid and subcloned into pEGFP eukaryotic expression vector. After the full-length sequence expected was confirmed by enzymatic digestion and sequencing,the human gastric carcinoma cell line SGC7901, which expressed a low level of VEGF-D, was transfected with the pEGFP/VEGF-D expression vector. Stable SGC7901 clones with high expression of VEGF-D were selected in vitro with G418, which were then combined and subcutaneously injected into nude mice to observe the density and morphology of lymphatic vessel. The outcomes were later compared with those of SGC7901 cells transfected with null vector(pEGFP) by immunostaining with a specific antibody LYVE-1.

RESULTSThe average weight of tumors in the pEGFP group (1.13+/-0.40) g at day 35, was significantly lower than that in the pEGFP/VEGF-D group (2.24+/-0.82)g (P<0.05). The lymphatic vessel density (LVD) in the pEGFP group (2.89+/-1.32) was significantly lower than that in the pEGFP/VEGF-D group (5.74+/-1.30)(P<0.01). There were dilated functional lymphatic vessels around the tumor margin.

CONCLUSIONVEGF-D may promote the growth and metastasis of tumor in gastric carcinoma by increasing the growth of lymphatic vessels.

Animals ; Cell Line, Tumor ; Humans ; Lymphatic Metastasis ; pathology ; Lymphatic Vessels ; pathology ; Mice ; Mice, Nude ; Stomach Neoplasms ; pathology ; Transfection ; Vascular Endothelial Growth Factor D ; genetics

Animals ; Borrelia burgdorferi ; genetics ; China ; DNA, Bacterial ; genetics ; isolation & purification ; Rodentia ; microbiology ; Sequence Analysis, DNA

OBJECTIVETo determine the effect of albumin administration on lung injury in traumatic/hemorrhagic shock (T/HS) rats.

METHODSForty-eight adult Sprague-Dawley rats were divided into three groups randomly (n=16 in each group): Group A, Group B, Group C. In Group A, rats underwent laparotomy without shock. In Group B, rats undergoing T/HS were resuscitated with their blood plus lactated Ringer's (twice the volume of shed blood). In Group C, rats undergoing T/HS were resuscitated with their shed blood plus additional 3 ml of 5% human albumin. The expression of polymorphonuclear neutrophils CD18/CD11b in jugular vein blood was evaluated. The main lung injury indexes (the activity of myeloperoxidase and lung injury score) were measured.

RESULTSSignificant differences of the expression of CD18/11b and the severity degree of lung injury were founded between the three groups. (P<0.05). The expression of CD18/CD11b and the main lung injury indexes in Group B and Group C increased significantly compared with those in Group A (P<0.05). At the same time, the expression of CD18/CD11b and the main lung injury indexes in Group C decreased dramatically, compared those in Group B (P<0.05).

CONCLUSIONSThe infusion of albumin during resuscitation period can protect lungs from injury and decrease the expression of CD18/CD11b in T/HS rats.

Albumins ; therapeutic use ; Animals ; CD11b Antigen ; metabolism ; CD18 Antigens ; metabolism ; Disease Models, Animal ; Neutrophils ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; drug therapy ; etiology ; metabolism ; Shock, Hemorrhagic ; complications ; metabolism ; Treatment Outcome ; Wounds and Injuries ; complications ; metabolism

Objective To monitor the co-infection status of Borrelia burgdorferi sensu lato (R.b.s.1) and spotted fever group Rickettsia (SFGR) in tourist areas of Heilongjiang province.Methods Polymerase chain reaction (PCR) was used to detect the 5S-23S rRNA intergenic spacer of B.b.s.1 and ompA of SFGR in ticks,dynamically collected from tourist areas of Heilongjiang province in 2010.Amplification products from positive ticks were sequenced,and phylogenetic analysis was conducted by Mega 5.0 software package.Results 849 ticks were collected from two tourist points,with the dominant ticks in Tiger Mountain and Jingpo Lake were Ixodes persulcatus and Haemaphysalis concinna.Regarding the Ixodes persulcatus from Tiger Mountain,the infection rates of B.b.s.1 and SFGR were 26.15％ and 10.05％.The infection rate of SFGR was 13.33％ in Haemaphysalis concinna and the B.b.s.1 was tndiscovered in the same ticks from Jingpo Lake.However,the co-infection could only be detected in Ixodes persulcatus of both tourist areas.Surveillance data showed that the major ticks were more likely to be appeared in July at Tiger Mountain and in June at Jingpo Lake.Data from the sequence analysis on B.b.s.1 showed that the B.b.s.1 in tourist areas could be classified into three different genotypes,other than B.garinii and B.afzelii.We first detected B.valaisiana-like group genotype in northeast of China.Results from the sequence analysis of SFGR positive products showed that the two DNA sequences of newly detected agents were completely the same as Rickettsia sp.HL-93 which was detected in Hulin and Rickettsia sp.H820 found in northeast,China.Conclusion The co-infection of B.b.s.1 and SFGR was detected in ticks from the tourist areas of Heilongjiang province,and data from the sequencing of specific fragment showed that various kinds of genotypes existed in this area.However; the rates of co-infectionitis-different according to environment,time and population that contributed to the kinds of and the index of ticks existed in the surveys points,also the infection rate of the ticks was studied.

Internal carotid arterial system is the main source of blood supply in brain. In forensic identi-fication practice, blunt injures of internal carotid arterial system usually cause complications such as throm-bus, aneurysm and arteriovenous fistula, etc. The deaths following delayed cerebral infarction or intracra-nial haemorrhage are not rare. At present, literature of deaths caused by blunt injuries of internal carotid arterial system mainly consist of case reports in China. This paper reviews related literature and case re-ports at home and abroad, and summarizes forensic medical features and identification method of the deaths caused by such injuries. The results show that blunt injures of internal carotid arterial system are related to the direct or indirect force on head and neck, which can result in exceed physiological range traction of head and neck, incision following basicranial fracture, etc. Such injuries are common in the cases as mechanical asphyxia, cervical manipulation, traffic accident and fall, etc. The artery should be examined carefully, and the relationship among injury, disease and death should be analysed correctly when no cause of infarction and hemorrhage was found in routine examination of such cases in forensic pratice. Because of the difficulty for exposing the artery completely in autopsy, angiography can be used to infer the location when necessary for improving the scientificity and reliability of the appraisal conclusion.

The medical insurance medical consumables were introuduced in terms of coding and standard implementation.A whole-process supervision method based on the codes of medical insurance medical consumbles was put forward to carry out catalog classification and selection,demand reporting and planned procurement,acceptance and storage management and use supervision,conditions monitoring and analysis and etc.The efficiency of various departments of clinical insitutitions was enhanced effectively for supervising clinical application of medical consumables,and the whole-process management of medical consumables was standardized.References were provided for the precision management of medical consumables.[Chinese Medical Equipment Journal,2023,44(9):74-77]

OBJECTIVETo observe the changes of serum free testosterone (FT) and testosterone secreting index (TSI) in ED patients, and to assess the contribution of these two indexes to the diagnosis of ED caused by endocrine factors.

METHODSWe studied 120 ED patients and 30 healthy men undergoing pre-marital medical check-up in Jiangsu Province Hospital of TCM by analyzing the scores on erectile function and desire domain in IIEF, testing the serum total testosterone, luteinizing hormone by chemiluminescent enzyme immunoassay (CLIA), measuring free testosterone by radioimmunoassay( RIA), and calculating TSI.

RESULTSOf the 120 ED patients, 5% and 1538% were below the reference norm of TT and FT values respectively. TT, FT and TSI decreased with age, with statistical with FT and TSI, but not with TT. FT and TSI statistically declined with lower IIEF score on ED domain, but this was not the case with TT. There were no significant differences in TI, FT and TSI among different sexual desire groups the ED patients.

CONCLUSIONFT is much more valuable than TF in the diagnosis of ED with hypogonadism. Both FT and TSI are important parameters in assessing the severity of ED.

Adult ; Aged ; China ; Erectile Dysfunction ; blood ; metabolism ; Humans ; Immunoenzyme Techniques ; methods ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Radioimmunoassay ; Surveys and Questionnaires ; Testosterone ; blood ; secretion

OBJECTIVETo establish experimental models for tumor neovascularization and to apply quantitative digital imaging analysis in the study.

METHODSAn endothelial tube formation model was established by human umbilical vein endothelial cells (HUVECs). A vasculogenic mimicry model was established by SGC-7901 gastric cancer cell line. Fertilized eggs were used to establish a chorioallantoic membrane angiogenesis model. Using gene transfection experiment, IRX1 tumor suppressor gene was chosen as a therapeutic target. Image Pro Plus (IPP) analysis software was used for digital vascular images analysis with parameters including points, lines, angles and integral absorbance (IA) for the tubular formation or vasculogenic mimicry.

RESULTSDigital image analysis by IPP showed that HUVEC tubular formation was significantly inhibited in IRX1 transfectant, compared with controls. The tubular numbers in three groups were 12.80 +/- 3.83, 29.00 +/- 5.34 and 28.20 +/- 4.32 (P<0.01). The connection points of tubules in three groups were 13.20 +/- 2.59, 25.00 +/- 2.24 and 24.60 +/- 3.21 (P<0.01). The tubular lengths of three groups were (821.5 +/- 12.5), (930.9 +/- 13.5) and (948.4 +/- 18.1) microm (P=0.022). The IA values of PAS stain in three groups were 3606 +/- 363, 14 200 +/- 1251 and 15 043 +/- 1220 (P<0.01). In chick chorioallantoic membrane model, the angular numbers of tubules in three groups were 6.41 +/- 2.60, 10.27 +/- 2.65 and 9.18 +/- 1.99 (P<0.01).

CONCLUSIONSThe endothelial tube formation model, vasculogenic mimicry model and chorioallantoic membrane angiogenesis model are useful for gene therapy and drug screening with targeting neoplastic vascularization. Professional image analysis software may greatly facilitate the quantitative analysis of tumor neovascularization.

Animals ; Cell Line, Tumor ; Cells, Cultured ; Chorioallantoic Membrane ; blood supply ; Diagnostic Imaging ; methods ; Homeodomain Proteins ; genetics ; metabolism ; physiology ; Human Umbilical Vein Endothelial Cells ; Humans ; Neovascularization, Pathologic ; Neovascularization, Physiologic ; Software ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transcription Factors ; genetics ; metabolism ; physiology ; Transfection