Evaluation of the Changes in microRNA Expression Profiles in Rat Primary Osteoblastic Cells Stimulated with Cotinine.

Objective: Smoking stands as a significant factor contributing to aberrations in bone metabolism, while microRNAs are intricately linked to the regulation of bone metabolism. This study aimed to identify cotinine-responsive microRNAs (miRNAs) and downstream regulatory pathways of their target genes involved in the regulation of osteoblastic cells, providing a foundation for new treatments targeting miRNAs for the bone metabolism imbalance induced by smoking. Methods: Primary osteoblastic cells of Sprague-Dawley rats were cultured through a modified enzymatic digestion method from the cranial bone of neonatal rats and stimulated with a high concentration of cotinine (1000 ng/mL) for 7 days. Then, miRNA gene chip technology was utilized to detect the changes in miRNA expression profiles in cotinine-stimulated osteoblastic cells, and differential expression profiles of cotinine-responsive miRNAs in osteoblastic cells were identified. Real-time polymerase chain reaction was used to detect the levels of significantly differentially expressed miRNAs in rat osteoblastic cells. Gene ontology (GO) and Kyoto encyclopedia of Genes and Genomes (KEGG) pathway analyses were utilized to predict target genes of these miRNAs to reveal the potential biological functions and pathways. Results: We identified 6 statistically differentially expressed miRNAs in the miRNA microarray analysis, of which 3 were upregulated and 3 were downregulated. We chose bone metabolism-related miRNAs as the miRNAs of interest. Quantitative real-time polymerase chain reaction was used to detect the expression levels of the differentially expressed miRNAs, and only miR-210 was significantly upregulated (3.34-fold), consistent with the microarray data. GO and KEGG pathway analyses of predicted miR-210 target genes revealed that miR-210 might participate in numerous signaling pathways, such as the RAS, Rap, PI3K-Akt, and calcium signaling pathways. Conclusion: We found that the strongly upregulated miR-210 may play an important regulatory role in osteoblast cells' biological behavior and bone formation function. The GO analysis results showed that miR-210 mainly involved protein binding, transporter activity, growth factor binding, and ion channel activity. According to the results of the KEGG analysis, miR-210 might negatively regulate the PI3K-Akt signaling pathway, thus affecting the proliferation of osteoblastic cells. These findings suggest that miR-210 may be a potential target for regulating the imbalance of bone metabolism caused by smoking, offering a new direction for clinical treatment of patients with bone metabolism-related diseases.

Survival and complications after neoadjuvant chemoradiotherapy versus neoadjuvant chemotherapy for locally advanced gastric cancer: a systematic review and meta-analysis.

Background: There is increasing evidence that neoadjuvant chemoradiotherapy is superior to neoadjuvant chemotherapy for patients with locally advanced gastric cancer. However, a number of studies have come to the opposite conclusion. Therefore, our meta-analysis is to evaluate the efficacy and safety of neoadjuvant chemoradiotherapy versus neoadjuvant chemotherapy in the treatment of locally advanced gastric cancer. Methods: We searched Wanfang Database, China National Knowledge Network database, VIP database, China Biomedical Literature Database, PubMed, Embase and Cochrane Library. The searched terms included'Stomach Neoplasms', 'Neoadjuvant Therapy' and 'Chemoradiotherapy'. The retrieval time was from the establishment of the corresponding database to September 2022, and our meta-analysis was performed using RevMan (version 5.3) and Stata (version 17) software. Results: A total of 17 literatures were included, which involved 7 randomized controlled trials and 10 retrospective studies, with a total of 6831 patients. The results of meta-analysis showed that compared with NACT group, the complete response rate(RR=1.95, 95%CI 1.39-2.73, p=0.0001), the partial response rate(RR=1.44, 95%CI 1.22-1.71, p=0.0001), the objective response rate(RR=1.37, 95%CI 1.27-1.54, p=0.00001), the pathologic complete response rate(RR=3.39, 95%CI 2.17-5.30, p=0.00001), the R0 resection rate(RR=1.18, 95%CI 1.09-1.29, p=0.0001) and 3-year overall survival rate(HR=0.89, 95%CI 0.82-0.96, p=0.002) of neoadjuvant chemoradiotherapy group were significantly improved. The results of subgroup analyses of gastric cancer subgroup and gastroesophageal junction cancer subgroup were consistent with the overall results. Meanwhile, the stable disease(RR=0.59, 95%CI:0.44-0.81, P=0.0010) of neoadjuvant chemoradiotherapy group was lower than that of neoadjuvant chemotherapy group, and there were no statistical significance in the progressive disease rate(RR=0.57, 95%CI:0.31-1.03, P=0.06), five-year overall survival rate(HR=1.03, 95%CI:0.99-1.07, P=0.839), postoperative complications and adverse reactions between the neoadjuvant chemoradiotherapy group and neoadjuvant chemotherapy group. Conclusion: Compared with neoadjuvant chemotherapy, neoadjuvant chemoradiotherapy might bring more survival benefits without significantly increasing adverse reactions. neoadjuvant chemoradiotherapy may be a recommended treatment for patients with locally advanced gastric cancer. Systematic Review Registration: https://inplasy.com/inplasy-2022-12-0068/, identifier INPLASY202212068.

LncRNA LINC00460 facilitates the proliferation and metastasis of renal cell carcinoma via PI3K/AKT signaling pathway.

Renal cell carcinoma (RCC) is one of the most prevalent cancers diseases in the worldwide. Long noncoding RNAs (LncRNAs) have been indicated as a mediator acted in tumorigenesis of RCC. LINC00460 has been reported to participate in many kinds of malignancies and promotes cancer progressions. However, the mechanism of LINC00460 on RCC is yet to be investigated. This study aimed to explore the potential function and regulation mechanism of LINC00460 in RCC. We analysed the LINC00460 expression and the prognosis in RCC patients using Gene Expression Profiling Interactive Analysis (GEPIA) and The Cancer Genome Atlas (TCGA) databases. LINC00460 level in normal renal cell line and RCC cell lines were examined by qRT-PCR. We study the effects of LINC00460 on proliferation, migration, invasion, apoptosis in RCC cells lines using a series of in vivo and in vitro experiments. RNA sequencing (RNA-seq) analysis was applied to searching potential LINC00460 related signal pathway in RCC. We identified the significant up-regulated expression of LINC00460 both in RCC tissues and cell. RCC patients with elevated LINC00460 expression have shorter survival. Up-expression of LINC00460 promoted cell proliferation, invasion and migration, meanwhile down-regulation of LINC00460 exerted inhibitory effect on these activities. We crucially identified that LNC00460 promotes development of RCC by influencing the PI3K/AKT pathway. Knockdown of LNC00460 decreased the phosphorylation of AKT and mTOR. The key finding of our study showed that LINC00460 functions as an oncogene in RCC pathogenesis by mediating the PI3K/AKT.

Survival and complications after neoadjuvant chemoradiotherapy versus neoadjuvant chemotherapy for esophageal squamous cell cancer: A meta-analysis.

OBJECTIVES: To compare the survival and complications of neoadjuvant chemoradiation (NCRT) versus neoadjuvant chemotherapy (NCT) for esophageal squamous cell carcinoma (ESCC). METHODS: We conducted a systematic literature search of the PubMed, Web of Science, Cochrane Library, EMBASE, CNKI, Wanfang Data, CBM, and VIP databases from inception to November 2021. Meta-analyses were performed using RevMan (version 5.3) and Stata version 15.0. RESULTS: A total of 18 studies were included, which involved 3137 patients, The results of the metaanalysis showed that the pathological complete remission rate (odds ratio [OR] = 5.21, 95% confidence interval [CI]: 2.85-9.50, p<0.00001) and complete tumor resection rate (OR = 2.31, 95% CI: 1.57-3.41, p<0.0001) in the NCRT group were significantly better than those in the NCT group. Our meta-analysis results showed that 1-, 3-, and 5-year survival rates (1-year overall survival [OS]: OR = 1.51, 95% CI: 1.11-2.05, p = 0.009; 3-year OS: OR = 1.73, 95% CI: 1.36-2.21, p<0.0001; 5-year OS: OR = 1.61, 95% CI: 1.30-1.99, p<0.00001) in the NCRT group were significantly higher than those in the NCT group. NCRT can lead a significant survival benefit compared with NCT and there was no significant difference between the two neoadjuvant treatments in terms of postoperative complications. CONCLUSION: The use of NCRT in the treatment of patients with ESCC patients showed significant advantages in terms of survival and safety relative to the use of NCT.

Androgen deprivation therapy and radiotherapy in intermediate-risk prostate cancer: A systematic review and meta-analysis.

Objectives: Androgen deprivation therapy combined with radiotherapy for intermediate-risk prostate cancer is still a matter of debate. We conducted a meta-analysis to evaluate the necessity of androgen deprivation therapy combined with radiotherapy for intermediate-risk prostate cancer patients. Methods: A comprehensive literature search of articles was performed in PubMed, Embase, Cochrane library, Web of Science, Chinese National Knowledge Infrastructure, Chinese Biological Medicine, Wanfang, and VIP Databases published between February 1988 and April 2022. Studies comparing the survival of patients diagnosed with intermediate-risk prostate cancer who were treated with androgen deprivation therapy combined with radiotherapy or radiotherapy alone were included. Data were extracted and analyzed with the RevMan software (version 5.3) and the Stata software (version 17). Results: Six randomized controlled trials and nine retrospective studies, including 6853 patients (2948 in androgen deprivation therapy combined with radiotherapy group and 3905 in radiotherapy alone group) were enrolled. Androgen deprivation therapy combined with radiotherapy did not provide an overall survival (HR 1.12, 95% CI 1.01-1.12, p=0.04) or biochemical recurrence-free survival (HR 1.23, 95% CI 1.09-1.39, P=0.001) advantage to intermediate-risk prostate cancer patients. Conclusion: Androgen deprivation therapy combined with radiotherapy did not show some advantages in terms of overall survival and biochemical recurrence-free survival and radiotherapy alone may be the effective therapy for intermediate-risk prostate cancer patients. Systematic review registration: https://inplasy.com/inplasy-2022-8-0095/, identifier 202280095.

Clinicopathologic Characteristics of Thyroid-like Low-grade Nasopharyngeal Papillary Adenocarcinoma: A Case Report.

Thyroid-like low-grade nasopharyngeal papillary adenocarcinoma (TL-LGNPPA) is an extremely rare neoplasm. It morphologically resembles papillary thyroid carcinoma and is immunochemically positive for thyroid transcription factor 1. Herein, we reported a case of TL-LGNPPA in a female patient of 35 years old. She felt pharyngeal foreign body sensation of unknown cause and subsequently developed dyspnea after activities. Nasopharyngeal plain computerized tomography showed no abnormalities, but laryngoscopy showed a peanut-sized smooth neoplasm with a pedicel at the posterior edge of the nasal septum. The mass was completely resected by nasopharyngoscopy. Histologic examination showed the tumor was composed of papillary configuration and tubular glands; each papilla was covered with cuboidal or columnar epithelial cells; tubular architecture and spindle cell component were also observed; some tumor cells had psammoma bodies. Immunohistochemically, tumor cells were positive for CKpan, CK7, CK19, VIM, and thyroid transcription factor 1, but negative for thyroglobulin, CK20, S-100, P63, P40, smooth muscle actin, CDX-2, and glial fibrillary acidic protein. This patient was diagnosed with TL-LGNPPA and followed up for 16 months, and metastasis and recurrence were not observed.

Effect of Different Titanium Surfaces on Maturation of Murine Bone Marrow-Derived Dendritic Cells.

Dendritic cells (DCs) play a pivotal role in the host response to implanted biomaterials. Osseointegration of titanium (Ti) implant is an immunological and inflammatory-driven process. However, the role of DCs in this complex process is largely unknown. This study aimed to investigate the effect of different Ti surfaces on DC maturation, and evaluate its subsequent potential on osteogenic differentiation of preosteoblasts. Murine bone marrow-derived DCs were seeded on Ti disks with different surface treatments, including pretreatment (PT), sandblasted/acid-etched (SLA) and modified SLA (modSLA) surface. Compared with DCs cultured on PT and SLA surfaces, the cells seeded on modSLA surface demonstrated a more round morphology with lower expression of CD86 and MHC-II, the DC maturation markers. Those cells also secreted high levels of anti-inflammatory cytokine IL-10 and TGF-ß. Notably, addition of conditioned medium (CM) from modSLA-induced DCs significantly increased the mRNA expression of Runx2 and ALP as well as ALP activity by murine preosteoblast MC3T3-E1 cells. Our data demonstrated that Ti disks with different surfaces lead to differential DCs responses. PT and SLA surfaces induce DCs mature, while DCs seeded on modSLA-Ti surface maintain an immature phenotype and exhibit a potential of promoting osteogenic differentiation of MC3T3-E1 cells.

Effects of Hypobaric Treatments on the Quality, Bioactive Compounds, and Antioxidant Activity of Tomato.

Hypobaric treatment is becoming a potential technology to protect fruits from postharvest decay. The objective of this study was to investigate the effects of hypobaric treatments on storage quality, bioactive compounds, and antioxidant activity of tomato fruit. In this study, green tomatoes (cv. "Fen guan") were treated with hypobaric pressures (0.04 and 0.07 MPa) at ambient temperature (20 ℃) for 28 d. The results showed that under hypobaric storage, the respiration rates significantly declined and the respiratory peaks postponed 12 and 8 d by 0.04 and 0.07 MPa treatments, respectively, compared to control. Total soluble solid, titratable acidity, ascorbic acid, and lycopene were retained by hypobaric treatment. Moreover, ascorbic acid contents treated with 0.04 and 0.07 MPa were, respectively, 37% and 26% higher than control at day 24 and the contents of total polyphenols were, respectively, 1.28 and 1.11 times higher than control. Production and accumulation of toxic substances were significantly restrained. The ethanol content decreased, respectively, by 53% and 84% than control. At later storage period, the superoxide dismutase activity in treated fruits was about 0.58 U/(g·FW·min), whereas only 0.29 U/(g·FW·min) in control. Hypobaric treatment not only maintained a high activity of superoxide dismutase and peroxidase (POD), but also improved antioxidant capacity. All the results indicated that hypobaric treatment was a potential helpful method to protect the quality and nutrition of tomato and prolong ripening of tomato. Furthermore, the effect of 0.04 MPa hypobaric treatment was found better than 0.07 MPa.

Characterization, promoter analysis and expression of the interleukin-6 gene in blunt snout bream, Megalobrama amblycephala.

Interleukin-6 (IL-6) is one of the most important multifunctional cytokines, playing essential roles in mediating the innate and adaptive immune responses. In this study, il-6 gene and its promoter from blunt snout bream, Megalobrama amblycephala, were characterized, and its expression at the transcript level in healthy fish and after bacterial infection was determined by quantitative real-time PCR. The results showed that the M. amblycephala il-6 (Mamil-6) cDNA had an ORF of 699 bp, encoding 232 amino acids, and contained 9 instable motifs in the 3' UTR. The deduced MamIL-6 possessed a 24-amino acid signal peptide and was located in the cytoplasm. Although sequence alignment and phylogenetic analysis revealed that IL-6 is poorly conserved in vertebrates, the protein and genomic structure of il-6 gene was well conserved. Analysis of the Mamil-6 promoter revealed the presence of a conserved TATA box and six major cis-regulatory elements, including C/EBPß (NF-IL6), AP-1, CRE, GRE, GATA and NF-κB binding sites. In healthy fish, Mamil-6 was the most abundant in the spleen. After Aeromonas hydrophila infection, Mamil-6 was significantly up-regulated in all 6 immune-related tissues examined, suggesting that Mamil-6 plays an important role in the blunt snout bream immune system.

Influence of Maxillary Sinus Width on Transcrestal Sinus Augmentation Outcomes: Radiographic Evaluation Based on Cone Beam CT.

BACKGROUND: Maxillary sinus elevation is a predictable procedure to vertically enhance bone volume in the posterior maxilla for successful implant placement. It is speculated that graft bone resorption and remodeling which require angiogenesis may be affected by the dimensions of maxillary sinus cavity. PURPOSE: The aim of this study is to investigate the effect of sinus width (SW) on the outcomes of transcrestal sinus lift with simultaneous implant placement based on cone beam CT (CBCT). MATERIALS AND METHODS: A total of 57 elevated sites in 33 patients were included in this study. All the patients were treated with transcrestal sinus lift procedure associated with simultaneous implant placement using a composite graft material of autogenous bone and Bio-Oss. For each patient, CBCT scans were performed preoperatively, immediately after surgery and 6 months after surgery. Measurements of the linear parameters were conducted on the preoperative and postoperative CBCT images. The correlation of SW with graft resorption (GR) was analyzed using Pearson's correlation test with or without the classification of residual bone height. RESULTS: The average width of maxillary sinus was 13.68 ± 2.66 mm. The mean height of apical graft bone decreased from 2.85 mm immediately after surgery to 1.38 mm after 6 months. A positive association between SW and GR (r = 0.323, p = .014) was found in general. CONCLUSION: The findings show that graft bone resorption in elevated sinus has a positive correlation with the SW.

Treatment of hyperoxic acute lung injury with stem cell transplantation in murine models: a brief review.

Hyperoxia has been a prominent clinical concern with the emergence of the intensive care unit and prolonged mechanical ventilation, along with the increasing use of hyperbaric oxygen therapy. Indeed, prolonged breathing of a high oxygen partial pressure may cause hyperoxic acute lung injury (HALI). To date, HALI has been a focus in the fields of pediatric and pulmonary medicine. However, no effective strategies have been developed for therapy for HALI due to the complicated mechanisms underlying the pathogenesis of HALI. In recent years, increasing studies have employed cell-based therapy for HALI. In this review, we summarize findings from available studies on therapy for HALI using stem cells in murine models and, based on concerns in this field, present our findings on cell-based therapy for HALI.

Anticancer Activity of Saponins from Allium chinense against the B16 Melanoma and 4T1 Breast Carcinoma Cell.

The cytotoxic substance of A. chinense saponins (ACSs) was isolated using ethanol extraction and purified with the D101 macroporous adsorption resin approach. We investigated the anticancer activity of ACSs in the B16 melanoma and 4T1 breast carcinoma cell lines. Methylthioninium chloride and hematoxylin-eosin staining with Giemsa dyestuff were used when the cells were treated with ACSs. The results showed that the cells morphologies changed significantly; ACSs induced cell death in B16 and 4T1 cells based on acridine orange/ethidium bromide double fluorescence staining, with the number and degree of apoptotic tumor cells increasing as ACS concentration increased. ACSs inhibited the proliferation of B16 and 4T1 cells in a dose-dependent manner. They also inhibited cell migration and colony formation and exhibited a concentration-dependent effect. In addition, ACSs apparently inhibited the growth of melanoma in vivo. The preliminary antitumor in vivo assay revealed that early medication positively affected tumor inhibition action and effectively protected the liver and spleen of C57 BL/6 mice from injury. This study provides evidence for the cytotoxicity of ACSs and a strong foundation for further research to establish the theoretical basis for cell death and help in the design and development of new anticancer drugs.

Structure Identification of Triterpene from the Mushroom Pleurotus eryngii with Inhibitory Effects Against Breast Cancer.

Breast cancer is the leading cause of death among women, with approximately 1 million diagnoses annually. Triterpenoids, which have cancer preventive or anti-tumour efficacy towards various tumour cells, may play a role in breast cancer prevention. In our previous study, an acetic ether (EtOAc) fraction from the sporocarp of the edible mushroom Pleurotus eryngii (P. eryngii) exhibited significant tumour cell growth inhibition both in vitro and in vivo. In this study, three pentacyclic triterpenoid compounds (1-3) were isolated from EtOAc extracts using chromatographic separation and were identified using nuclear magnetic resonance (NMR) and mass spectrometry (MS). The compounds were 2, 3, 6, 23-tetrahydroxy-urs-12-en-28 oic acid (1), 2,3,23-trihydroxy-urs-12-en-28 oic acid (2) and lupeol (3). All three purified triterpenes showed significant inhibitory activity against breast cancer MCF-7 cell lines in vitro, with the greatest activity exhibited by compound 1, followed by compound 2 and 3. The IC(50) values were 15.71, 48 and 66.89 µM, respectively. Our study may help elucidate the health benefits of P. eryngii mushroom consumption.

[Screening and antibacterial function of Bacillus amyloliquefaciens X030].

OBJECTIVE: We isolated 339 bacillus strains from 72 soil samples all over the country, then purified their antimicrobial compounds and studied the antibacterial activity, to enrich bacillus resources and explore their second metabolites. METHODS: A bacillus strain with strong antibacterial activity was selected by dilution plate and water bath heating from a soil sample from a peanut plantation in Henan Province; this strain was identified according to morphological observation, physiological and biochemical characteristics, and consequences of 16S rRNA homologous analysis. Antibacterial compound from the identified strain, Bacillus amyloliquefaciens X030, was separated and purified by acetone precipitation, Sephadex chromatography, C18 reverse phase column chromatography. Its molecular weight was analyzed by LC-MS/MS. The antibacterial activity was characterized by disc diffusion and plate two-way cultivation. RESULTS: Bacillus amyloliquefaciens was isolated that not only has antibacterial activity against Staphylococcus aureus, Candida albican and Saccharomycetes; but also against Pyriculariaoryzae, Chili pointed cell anthrax, Gloeosporium eriobotryae speg and Phytophthora parasitica. The compound was confirmed as polypeptide. CONCLUSION: Bacillus amyloliquefaciens X030 can produce a polypeptide that inhibits pathogenic bacteria and plant pathogenic fungi.

Culture and identification of human bone marrow mesenchymal stem cells from alveolar ridge dental implant site.

OBJECTIVE: The objective of this study was to establish the culture method of human bone marrow mesenchymal stem cells (BMSCs) from alveolar bone marrow. METHODS: Alveolar bone marrow complex samples were obtained from 35 patients, 22 to 65 years of age, during the course of dental implant treatment by low-speed method. Bone marrow mesenchymal stem cells were seeded and maintained in culture with 10% fetal bovine serum. The form of the cultured cells was observed under inverted microscope, and the cell proliferation capacity was detected. Cell cycle and the antigen expression of P3 BMSCs were measured with flow cytometry. RESULTS: From a small volume (about 0.1-0.2 mL) of bone marrow complex, alveolar BMSCs expanded at a success ratio of 29 (83%) of 35. There is dysmorphism in cultured cells, which mainly were long spindle, polygon, and triangle. Flow cytometry instrument test showed that the cells in G0/G1 phase were an average of 79.29% ± 1.70% and in S phase were an average of 11.09% ± 0.87%. For antibody expression rate: CD29 is 88.00% ± 1.56%, CD44 is 88.15% ± 1.64%, CD34 is 0.42% ± 0.10%, CD45 is 0.45% ± 0.12%, and CD11b is 0.45% ± 0.14%. CONCLUSION: The cells of the marrow complex obtained by low-speed method in implant site preparation cultured in vitro were identified as BMSCs through the morphological observation and the flow cytometry. It is a kind of feasible and simple culture method of human primary BMSCs.

Development of TaqMan-based qPCR method for detection of caprine arthritis-encephalitis virus (CAEV) infection.

A specific and sensitive two-step TaqMan real-time PCR has been developed for rapid diagnosis of caprine arthritis-encephalitis virus (CAEV) infection by using a set of specific primers and a TaqMan probe targeting a highly conserved region within the gene encoding the viral capsid protein (CA). The assay successfully detected CAEV proviral DNA in total DNA extracts originating from cell culture, whole blood samples and isolated PBMCs, with a lower detection limit of 10(2) copies and a linear dynamic range of 10(5) to 10(10) copies/ml. There was no cross-reaction with other animal viruses (e.g., goat pox virus, bovine leukemia virus, bovine mucosal disease virus, swine influenza virus and Nipah virus). When applied in parallel with serological AGID and conventional PCR for detection of CAEV in field samples, this assay exhibited a higher sensitivity than these traditional methods, and 7.8 % of the 308 specimens collected in the Shanxi and Tianjin regions of China from 1993 to 2011 were found to be positive. Thus, the TaqMan qPCR assay provides a fast, specific and sensitive means for detecting CAEV proviral DNA in goat specimens and should be useful for large-scale detection in eradication programs and epidemiological studies.

The binding of CpG-oligodeoxynucleotides to cell-surface and its immunostimulatory activity are modulated by extracellular acidic pH.

Both the binding of CpG-oligodeoxynucleotides (CpG-ODNs) to cell-surface and its immunostimulatory activity were modulated by extracellular pH in present study. At neutral pH (pH 7.4), the binding of CpG-ODN to splenocyte-surface, as well as that of non-CpG-ODN, was competitively inhibited by non-specific DNA-Herring sperm DNA in a dose dependent manner, indicating their binding sites have no specificity for CpG-motif. When the extracellular pH shifted to acidic (pH 6.4), however, their binding to cell-surface markedly increased, and only the binding of non-CpG-ODN instead of CpG-ODN was inhibited by Herring sperm DNA, implying such pH change enabled CpG-ODN bind to its specific binding-site. Consistently, lymphocytes appeared more sensitive to the stimulation of CpG-ODN at acidic pH, and Herring sperm DNA inhibited the CpG-ODN-induced TNF production from splenocytes at pH 7.4, but not at pH 6.4. These results suggest the existence of membrane receptor that specifically engages CpG-ODN with high affinity only at acidic pH, and support the hypothesis that the binding CpG-ODN to its specific membrane receptor and subsequently triggering of CpG-related signaling occurred within acidified endosomes.

Molecular Cloning of a Gene Suppressed by Heat Shock and Its Expression Characteristics.

On the base of a cDNA fragment cloned from a human osteosarcoma cell line (HOS-8603) which was taken as the model of heat shock by DDRT-PCR, the cDNA library was screened by the probe made from this fragment, and the colony containing full length cDNA of this cDNA fragment (HSSG-1) was identified. Sequencing data indicated that this cDNA consisted of 1 456 bp, coding for 276 amino acids. Computer aided homology analysis did not find the same sequence published. The result of RNA dot hybridization suggested that HSSG-1 was expressing in many kinds of organs and cells, and the down regulating of its expression level caused by heat shock was general. Therefore, this may be a gene that is suppressed by heat shock.