OBJECTIVETo investigate the injury stress responses caused by ischemia reperfusion and its effects on the salivary secretory function of rat submandibular glands.

METHODSAn in situ ischemia reperfusion experimental model of rat submandibular glands was developed. The rat submandibular glands were subjected to 90 min of ischemia without denervation followed by reperfusion for 1, 12, 24, and 72 h. Salivary secretion, histological changes, reactive oxygen species (ROS) levels, and cellular apoptosis of the involved submandibular glands were detected after reperfusion.

RESULTSThe secretory function of the glands decreased at 1 and 12 h, and the saliva secretion gradually had the same value as that of the control sample 72 h after reperfusion. Increasing inflammatory cells infiltration, cellular atrophy, and tissue edema were observed especially after reperfusion for 12 h. The level of ROS and the number of apoptotic cells exhibited the same tendency, and higher ROS levels and more apoptosis cells 1 and 12 h after reperfusion were observed.

CONCLUSIONOur study suggests that ischemia reperfusion can cause a series of injury stress responses in submandibular glands, which might have an important function in the early phase dysfunction of transplanted submandibular glands.

Animals ; Apoptosis ; Rats ; Reperfusion Injury ; Submandibular Gland

BACKGROUND: Cryopreservation can decrease tissue and organ immunogenicity. The effects of cryopreservation on cell immunogenicity are disputed.OBJECTIVE: To investigate the effects of liquid nitrogen cryopreservation on osteoblast immunogenicity. DESIGN: Randomized,controlled ,paired-sample experiment. SETTING: This study was performed in the Laboratory Center, Qilu Hospital Affiliated to Shandong University between July 2003 and March 2004. MATERIALS: Four New Zealand rabbits of either gender were included for this study. 3H-TdR was provided by Nuclear Medicine Institute of Shandong University. METHODS: Osteoblasts were cultured from the tibial periosteum of New Zealand rabbits and then cryopreservated in the liquid nitrogen for 3 months and defrosted. Cryopreservated and thawn osteoblasts were set as cryopreserved group and freshly cultured osteoblasts were set as non-cyropreserved group. Major histocompatibility complex (MHC)-I positive rate was examined by flow cytometry assay prior to and after cryopreservation. Simultaneously, mixed lymphocyte-osteoblast cultures were established. Lymphocyte stimulation index was calculated after counting the flares using β liquid scintilloscope. MAIN OUTCOME MEASURES: MHI-I antigen positive rate and lymphocyte stimulation index prior to and after cryopreservation of osteoblasts. RESULTS: MHI-I antigen positive rate and lymphocyte stimulation index of osteoblasts was significantly higher in the non-cryopreserved group than in the cryopreserved group (P＜0.01). CONCLUSION: The immunogenicity of cryopreserved osteoblasts was significantly decreased. Liquid nitrogen cryopreservation is an ideal method to decrease the immunogenicity of osteoblasts.

Objective: To provide an efficeut protocol for constructing recombinant adenovirus, an in vitro ligation was used instead of homologous recombination. Methods: Gene BMP-2 was ligated into pShuttle 2 vector ( pShuttle 2-BMP-2 ) and then fragment containing BMP-2 gene and promoter pcmvie excised by PI-SCe Ⅰ and Ⅰ-Ceu Ⅰ endonuclease. The fragment was further combined with adenovirus vector (Adeno-X-BMP-2) , which was finally linearized with Pac Ⅰ and trans-fered to HEK293 to package adenovirus particles. Results: Both PCR assay and restiction analysis showed that the recombined rectors pShuttle2-BMP-2 and Adeno-X-BMP-2 contains the target BMP-2 gene. THe packaged adenovirus was also i-dentified by PCR assay with specific primers for BMP-2. Conclusions: The BMP-2 incorporated recombinant adenovirus was obtained and this laid a foundation for further study on BMP-2 mediated gene therapy. The in vitro ligation method de-scinbed here for constructing recombined adenovirus was more efficient than traditional homologous recombination.

Hepatitis B virus (HBV) can cause a variety of hepatitis B-related diseases in infected people, and there is no specific drug for treatment. China, with a large population base, is the country with the heaviest burden of HBV infection in the world. Therefore, hepatitis B vaccination is particularly important in the prevention and control of this disease. However, some vaccinees did not produce effective protective immune response after vaccination according to the recommended hepatitis B vaccine immunization schedule. The purpose of this review is to analyze the influential factors of non/low immune response after hepatitis B vaccination from the aspects of organism and vaccine, and explore the mechanism of non/low immune response, so as to provide scientific basis for the prevention and treatment of hepatitis B in China.

OBJECTIVETo investigate the influence of carbon monoxide on the expression of adhesion molecules stimulated by inflammatory cytokines on human gingival fibroblasts.

METHODSHuman gingival fibroblasts were stimulated with 50 ng x mL(-1) tumor necrosis factor (TNF)-alpha and 10 ng x mL(-1) interleukin (IL)-1beta concurrently in the presence or absence of 500 micromol x L(-1) carbon monoxide releasing molecule (CORM). Expression of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1 at protein and mRNA level was examined by Western blot and reverse transcription polymerase chain reaction (RT-PCR), respectively. Activity of transcription factor NF-kappaB was evaluated by reporter gene assay.

RESULTSExpression of ICAM-1 and VCAM-1 on human gingival fibroblasts increased dramatically after concurrent stimulation of TNF-alpha and IL-1beta, while CORM inhibited the upregulation of ICAM-1 and VCAM-1. CORM decreased the activity of NF-KB stimulated by TNF-alpha and IL-1beta.

CONCLUSIONCarbon monoxide could be a promising way in treating of periodontitis.

Carbon Monoxide ; Cells, Cultured ; Fibroblasts ; Gingiva ; Humans ; Intercellular Adhesion Molecule-1 ; Interleukin-1beta ; NF-kappa B ; RNA, Messenger ; Tumor Necrosis Factor-alpha ; Vascular Cell Adhesion Molecule-1

Objective To investigate the injury stress responses caused by ischemia reperfusion and its effects on the salivary secretory function of rat submandibular glands. Methods An in situ ischemia reperfusion experimental model of rat submandibular glands was developed. The rat submandibular glands were subjected to 90 min of ischemia without denervation followed by reperfusion for 1, 12, 24, and 72 h. Salivary secretion, histological changes, reactive oxygen species (ROS) levels, and cellular apoptosis of the involved submandibular glands were detected after reperfusion. Results The secretory function of the glands decreased at 1 and 12 h, and the saliva secretion gradually had the same value as that of the control sample 72 h after reperfusion. Increasing inflammatory cells infiltration, cellular atrophy, and tissue edema were observed especially after reperfusion for 12 h. The level of ROS and the number of apoptotic cells exhibited the same tendency, and higher ROS levels and more apoptosis cells 1 and 12 h after reperfusion were observed. Conclusion Our study suggests that ischemia reperfusion can cause a series of injury stress responses in submandibular glands, which might have an important function in the early phase dysfunction of transplanted submandibular glands.

A large-scale vaccination of coronavirus disease-19 (COVID-19) in adults has been conducted for nearly a year, and there is a growing recognition that immunization for children is also essential. It has been months since emergency use of pediatric COVID-19 vaccine was approved, we reviewed the prevalence and transmission of COVID-19 in children. The prevalence of COVID-19 in children is reduced due to vaccination even in a Delta prevalent period, so an increase in the vaccination rate is needed in children. Although the precise role of children in the transmission requires more research to uncover, they likely played a significant role, according to the available literature. We also described four candidate COVID-19 vaccines for children on their safety and immunogenicity and the impact of severe acute respiratory syndrome coronavirus 2 variants on childhood vaccination. Safety issues on pediatric vaccines post-approval, like adverse events following immunization and adverse events of special interest require studies on long-term and effective regulatory mechanisms.

OBJECTIVETo establish a novel nude mice model which can be visualized in real time and detected in a continuous and dynamic way for the development and metastasis of adenoid cystic carcinoma.

METHODSHuman adenoid cystic carcinoma cells, ACCM cell line, were infected with retroviral vector of pLEGFP-N1 and then screened for a single colony of ACCM-GFP cells. Cell proliferation and morphological analysis were conducted for ACCM and ACCM-GFP cells. Nude mice lingual carcinoma model was set up with ACCM-GFP cells injection and real time observation with fluorescence imaging on ACCM-GFP tumors was performed subsequently. Histological assay was analyzed for ACCM and ACCM-GFP tumors as well.

RESULTSACCM-GFP cells were able to express GFP stably in the long term. ACCM and ACCM-GFP cells showed no significant difference in cell proliferation and morphology, and no significant difference of histological characteristics in vivo could be found between ACCM and ACCM-GFP tumors. Tumor development could be monitored in real time with fluorescence imaging system in vivo.

CONCLUSIONGFP-expressing ACCM tumor model can be applied to detect and observe its development in the long term in a noninvasive, real time and dynamic way. It is also a kind of ideal in vivo mouse model for adenoid cystic carcinoma research.

Animals ; Carcinoma, Adenoid Cystic ; Cell Line, Tumor ; Cell Proliferation ; Disease Models, Animal ; Green Fluorescent Proteins ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude

OBJECTIVETo understand the distributions of DNA and neutralizing antibodies of human papillomavirus (HPV)16, 18 in 18-45 year-old women.

METHODSTotally, 1494 women were enrolled through multistage random sampling in Funing, Jiangsu province. Cervical exfoliated cells were collected from them for HPV DNA testing, and serum samples were taken from them for the detection of HPV16, 18 neutralizing antibodies by using pseudovirion-based neutralization assay(PBNA).

RESULTSAmong the 1494 women, 28(1.9%) and 188(12.6%) were positive for DNA and neutralizing antibody of HPV16 respectively, and 15(1.0%) and 60(4.0%) were positive for DNA and neutralizing antibody of HPV18, respectively. There were no significant differences in the detection rates of DNA and neutralizing antibody of HPV16, 18 among different age groups. About 16.7% of the women were infected with HPV16, 18, or both.

CONCLUSIONIn Funing county of Jiangsu province, most women aged 18-45 years has no immunity to HPV16 and 18, indicating that they are appropriate targets for HPV 16/18 vaccination.

Adolescent ; Adult ; Antibodies, Neutralizing ; isolation & purification ; Antibodies, Viral ; isolation & purification ; China ; DNA, Viral ; isolation & purification ; Female ; Human papillomavirus 16 ; immunology ; Human papillomavirus 18 ; immunology ; Humans ; Middle Aged ; Papillomavirus Infections ; prevention & control ; Papillomavirus Vaccines ; Young Adult

CRISPR-Associated Protein 9 ; Gene Editing ; CRISPR-Cas Systems/genetics*