Phylogenetic analysis of the Mongolian gerbil (Meriones unguiculatus) from China based on mitochondrial genome.

Meriones unguiculatus (Gerbillinae, Rodentia) is widely used as an animal model of human disease. Here, we provide the first report of the complete mitochondrial genome sequence of M. unguiculatus (GenBank accession Nos. KF425526 and NC_023263). The sequence contained the conserved vertebrate pattern of 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and 1 major noncoding region. We identified one extended termination-associated sequence and one conserved sequence block in the non-coding region. The putative origin of replication for the light strand (OL) was 35 bp long. The OL stem and adjacent sequences were highly conserved, but the loop region differed from those of other rodent species. Base composition and codon usage of the 13 protein-coding genes in M. unguiculatus were compared with those of 23 rodent species with previously sequenced mitochondrial genomes. An A+T content of 63.0% was present in M. unguiculatus; this is similar to the Murinae average (62.4 ± 0.8%) and falls between the average for Mus musculus (63.1 ± 0.1%) and Rattus sp (61.7 ± 0.4%). The AT and GC skew values of M. unguiculatus were 0.035 and -0.28, respectively, similar to those of Cricetinae species (0.057 ± 0.05 and -0.31 ± 0.05). The codon families exhibited similar abundance in all 24 species. Analysis of phylogenetic relationships with 23 other rodent species using neighbor-joining and maximum likelihood protocols and the 12 protein-coding regions on the H strand showed that M. unguiculatus should be classified as genus Meriones, sub-family Gerbillinae, family Muridae.

Expression of EpCAM and Wnt/ ß-catenin in human colon cancer.

The aims of this study were to explore the correlation between the expression of EpCAM and the Wnt/ß-catenin pathway in human colon cancer and its clinical significance for the evaluation of cancer prognosis. Samples from colon cancer, para-carcinoma, or benign intestinal tissue from individual patients (50) and from normal intestinal mucosal tissues (20) were obtained from the Pathology Department of the Shandong Province Binzhou People's Hospital (Shandong, China). Immunohistochemistry was used to detect the expression levels of EpCAM and ß-catenin proteins in these tissues, and the prognoses of the patients from whom the samples were derived were determined on follow-up examination. The corresponding in vitro mechanistic siRNA experiments were subsequently performed in the human colon cancer cell line HCT116 to observe the regulatory effects of silencing EpCAM expression on the Wnt/ß-catenin pathway. From these analyses, we determined that the expression levels of EpCAM and ß-catenin were higher in cancer tissues compared with other tissues from the same patient, and that the expression of EpCAM and Wnt/ß- catenin in colon cancers were positively correlated. The prognostic analysis showed an inverse correlation between EpCAM and Wnt/ß- catenin expression and patient prognosis. A further examination of cellular mechanisms confirmed that the silencing of EpCAM led to decreased expression of Wnt/ß-catenin, and thus reduced proliferation and increased the apoptosis ratio in the cells. These results suggest that suppression of EpCAM might be a new approach for treating colon cancer.

Kaposi's sarcoma-associated human herpesvirus-8 encodes homologues of macrophage inflammatory protein-1 and interleukin-6.

Human herpesvirus-8 (HHV-8) has been detected in Kaposi's sarcoma (KS) lesions of all types (AIDS-related, classical and endemic), in body-cavity-based B-cell lymphomas (BCBLs) and in lesions of multicentric Castleman's disease (MCD). We have identified a major gamma-herpesvirus-divergent locus (DL-B) in HHV-8 DNA encoding several HHV-8 unique open reading frames (ORFs), including a homologue of interleukin-6 (IL-6) and two homologues of macrophage inflammatory protein MIP-1. We show that the HHV-8-encoded IL-6 homologue (vIL-6) shares functional properties with endogenous IL-6 proteins and that both vIL-6 and vMIP-1 transcripts are present at high levels following butyrate induction of an HHV-8' BCBL cell line. Low amounts of constitutive vIL-6, but not vMIP-1, mRNA were also detected. The presence of a functional IL-6 homologue encoded by HHV-8 may provide a mechanistic model for the hypothesized role of HHV-8 in KS, MCD and BCBL that involves the mitogenic effects of vIL-6 on surrounding cells. MIP-1 proteins may enhance these effects through the chemotactic recruitment of endogenous cytokine-producing cells into affected tissues and could potentially influence HIV disease progression in coinfected individuals through interactions with the HIV co-receptor CCR-5.

[Comparison of clinical characteristics of patients with different types of laryngopharyngeal reflux disease].

Objective: To compare the clinical characteristics of patients with different type of laryngopharyngeal reflux disease in order to study the effect of non-acid reflux on laryngopharyngeal reflux disease. Methods: From January 2015 to January 2020, 349 inpatients or outpatients suspected of having laryngopharyngeal reflux underwent 24-hour multichannel intraluminal impedance pH monitoring (MII-pH). There were 303 male and 46 female patients, with an average age of 56.03 years old ranged from 25 to 81 years old. The reflux symptom index (RSI)and reflux findings score(RFS)were recorded before MII-pH monitoring. The number of acid reflux events and non-acid reflux events in hypopharynx were counted. It was defined mainly acid reflux type when the ratio of acid reflux to all reflux events was greater than 50%, mainly non-acid reflux type when the ratio of non-acid reflux to all reflux events was greater than 50%. The clinical characteristics of patients with different type of reflux were compared. SPSS 19.0 software was used for statistical analysis, and multiple independent samples were compared between groups. The quantitative data were analyzed by multivariate analysis of variance, and the counting data were analyzed by chi-square test, the difference was statistically significant when P<0.05. Results: The 24-hour MII-pH showed that there were 90 patients with no reflux events, 51 patients with mainly acid reflux type, 198 patients with mainly non-acid reflux type and 10 patients with equal acid reflux events and non-acid reflux events. Statistics showed that the RSI(10.72±4.40), RFS(7.70±2.73) and the average number of reflux events(0) in the group without reflux events were significantly lower than those in patients with mainly acid reflux type (RSI 13.16±6.62,RFS 10.08±3.03,average number of reflux events 5.33±3.15,P<0.05) and mainly non-acid reflux type(RSI 13.25±5.54,RFS 8.81±2.54,average number of reflux events 7.93±5.26, P<0.05). There was no significant difference in RSI between the mainly non-acid reflux type group and the mainly acid reflux type group, but the RFS of the mainly non-acid reflux type group was significantly lower than that of the mainly acid reflux type group. The average number of reflux events in the mainly non-acid reflux group was significantly higher than that in the mainly acid reflux type group (P<0.05). Conclusion: The results show that non-acid reflux plays a certain role in laryngopharyngeal reflux disease, but the effect of acid reflux is greater.

[Therapeutic effects of endoscopic posterior cordotomy for bilateral vocal cord paralysis].

Objective: To evaluate the efficacy of the endoscopic bilateral posterior transverse partial cordotomy in patients with upper airway obstruction due to bilateral vocal fold paralysis. Methods: A retrospective analysis of 48 cases of upper airway obstruction due to bilateral vocal fold paralysis, who were admitted to Department of Otolaryngology Head and Neck Surgery, the Sixth Medical Center of Chinese PLA General Hospital from July 2009 to July 2019, was performed, including 13 males and 35 females. Patients' ages ranged from 27 to 83 years old. All patients underwent bilateral vocal fold posterior resection. Results: Among the 48 patients, 1 patient was lost to follow-up, and the remaining 47 patients were followed up for 5 months to 10 years . None of the 47 patients had a recurrence or severe complications. 89.58% (43/48) patients reconstructed a reliable and effective airway and 88.89% (40/45) patients were decannulated in 1-3 months postoperatively, with the median decannulation time of 1 month. Recovery rate of swallowing function and satisfactory pronunciation were 97.92% (47/48) and 95.35% (41/43) respectively. Conclusions: Endoscopic bilateral posterior transverse partial cordotomy can establish a reliable and effective airway and maximize the protection of swallowing and voice functions. At the same time, it is a safe, reliable, simple and minimally invasive treatment option.

Novel viral sequences related to human T-cell leukemia virus in T cells of a seropositive baboon.

Antibodies reactive with proteins of human T-cell leukemia virus (HTLV) can be found in Old World monkeys. A T-lymphocyte cell line established from a seropositive baboon (Papio cynocephalus) was analyzed for the presence of viral DNA sequences. The provirus found in these cells was related to but distinct from HTLV subgroup I. These results add to recent evidence from human studies that HTLV represents a spectrum of infectious T-lymphotropic retroviruses that includes closely and distantly related members.

Transformation and cytopathogenic effect in an immune human T-cell clone infected by HTLV-I.

Human T-cell leukemia-lymphoma virus (HTLV) is a human C-type retrovirus that can transform T lymphocytes in vitro and is associated with certain T-cell neoplasms. Recent data suggest that, in the United States, patients with acquired immunodeficiency syndrome (AIDS), homosexual men with lymphadenopathy, and hemophiliacs have had significant exposure rates to HTLV, whereas matched and unmatched control American subjects have rarely been exposed to this agent. In the present experiments, T cells specifically reactive against HTLV were propagated from a patient whose HTLV-bearing lymphoma was in remission. The T cells were cloned in the presence of the virus and an HTLV-specific cytotoxic T-cell clone was isolated. This clone was infected and transformed by the virus, with one copy of an HTLV-I provirus being integrated into the genome. This T-cell clone did not exhibit the normal dependence on T-cell growth factor (interleukin-2) and proliferated spontaneously in vitro. Exposure of the clone to HTLV-bearing, autologous tumor cells specifically inhibited its proliferation and resulted in its death. These results may have implications for HTLV-associated inhibition of T-cell responses.

Functional properties of antigen-specific T cells infected by human T-cell leukemia-lymphoma virus (HTLV-I).

Tetanus-toxoid specific helper-inducer T-cell clones, which had been infected and transformed by human T-cell leukemia-lymphoma virus (HTLV-I), were obtained from an antigen-specific human T cell line by using a limiting dilution technique in the presence of the virus. These HTLV-I-infected T-cell clones proliferated specifically in response to soluble tetanus toxoid but, unlike normal T cells, they could do so in the absence of accessory cells. The HTLV-I-infected T-cell clones did not present the antigen to autologous antigen-specific T cells that were not infected with HTLV-I. The capacity of helper-inducer T cells to retain antigen-specific reactivity after infection by HTLV-I, while losing the normal T-cell requirement for accessory cells, has clinical and theoretical implications.

Alterations in cytotoxic and helper T cell function after infection of T cell clones with human T cell leukemia virus, type I.

HTLV-I is a transforming human retrovirus that is an etiologic agent of adult T cell leukemia/lymphoma. To investigate the effects of this virus on T cell functions, two OKT3+, OKT4+, OKT8- cytotoxic clones (8.7 and 8.8) specific for allogeneic cells bearing DPw2, a class II histocompatibility antigen, were studied before and after infection with HTLV-I. The clones retained cytotoxic function for up to 70 d after exposure to HTLV-I, even without subsequent antigenic stimulation, but then lost their cytotoxic activity. Prior to infection with HTLV-I, clone 8.8 also lysed OKT3 hybridoma cells; after infection, cytotoxic activity against these OKT3-antibody bearing cells was lost in parallel with the loss of activity against DPw2-bearing target cells. In addition, expression of T3 surface antigen by HTLV-I-infected 8.8 cells was decreased at a time when they lost their cytotoxic activity, possibly contributing to the loss of cytotoxic function. Finally, clone 8.8 could provide help for nonspecific IgG production by autologous B cells when stimulated with irradiated DPw2-bearing non-T cells. After infection with HTLV-I, this helper function became independent of DPw2-stimulation and persisted even when the cytotoxic activity was lost. An OKT4+ T cell clone thus could simultaneously manifest both cytotoxic and helper T cell activities, and these activities were differentially affected after HTLV-I infection.

Infection of human T lymphotropic virus-I-specific immune T cell clones by human T lymphotropic virus-I.

Human T lymphotropic virus-I (HTLV-I)-specific T cell lines were established and cloned. K5, an OKT8+ clone bearing multiple proviral integration sites, retained its HTLV-I-specific cytotoxicity and a normal dependence on interleukin 2 (IL-2), indicating that there is a finite number of transforming integration sites. R2, an OKT4+ HTLV-I-infected clone, initially mounted a proliferative response to HTLV-I; but then its IL-2-independent proliferation increased and the antigen specificity was lost. All HTLV-I-infected clones tested including K7, another OKT8+ transformed cytotoxic clone that had lost its reactivity, expressed comparable levels of T cell receptor beta-chain (TCR-beta) messenger (m)RNA. Although clones K5 and K7 had different functional properties, they had the same rearrangement of the TCR-beta gene, suggesting that they had the same clonal origin. These data indicate that HTLV-I-specific T cells retain their immune reactivity for variable periods of time following infection, but then usually lose it; in some cases, however, no alteration in function can be detected. The data also suggest that different consequences can take place in the same clone depending on the pattern of retroviral infection.

[The effect of regional neck dissection on positive cervical lymph node of cN0 laryngeal carcinoma].

Objective:To evaluate the efficacy and safety of regional neck dissection in the treatment of cN0 laryngeal carcinoma with positive cervical lymph node. Method:A retrospective analysis of 120 cases with cN0 laryngeal squamous cell carcinoma who received the first time for primary tumor resection and regional neck dissection (Ⅱ-Ⅳ) in our hospital during the period of 2000.01-2016.06 were performed. Twenty-two patients with lymph node positive (pN+) were selected by postoperative paraffin pathology in Ⅱ-Ⅳ region and followed up to 2017.06. The recurrence rate, survival rate and survival related regression analysis of patients with stage cN0 pN+ laryngeal carcinoma were analyzed. Result:The cN0 laryngeal cancer occult metastasis rate was 18.33% (22/120) in regional neck dissection. Local recurrence, regional recurrence, distant metastasis rates of 3 and 5 years were 41.18%, 17.65%, 17.65% and 40.00%, 13.33%, 20.00%, respectively in cN0 pN+ patients. The overall survival rates of 3 and 5 years were 61.2% and 30.6% respectively, and the disease-free survival rate was 31.8% and 22.7%. There was no significant difference in overall and disease-free survival between the T staging or clinical classification (P>0.05). Cox regression analysis showed that overall survival was related to age and local-regional (RR=11.421, P=0.001, RR=5.211, P=0.022). Logistic multivariate regression analysis showed that local-regional recurrence was not related to each factor (P>0.05). Conclusion:Local recurrence rate and mortality rate of cN0 pN+ laryngeal carcinoma are higher, survival rate is lower, however, neck recurrence rate is low.Therefore, Ⅱ-Ⅳ neck dissection is a safe and effective treatment for neck of cN0 pN+ laryngeal carcinoma.

Expression of human herpesvirus 8-encoded cyclin D in Kaposi's sarcoma spindle cells.

BACKGROUND: Human herpesvirus 8 (HHV-8) DNA sequences have been detected in Kaposi's sarcoma, in primary effusion lymphoma (an unusual high-grade non-Hodgkin's lymphoma seen primarily in patients with acquired immunodeficiency syndrome [AIDS]), and in Castleman's disease (a rare lymphoproliferative disorder); however, proof that HHV-8 is involved in the pathogenesis of these diseases remains to be established. HHV-8 contains a gene, i.e., v-cyclin D, that is a homologue of the cellular cyclin D2 gene, which encodes a protein that promotes passage through G1 phase of the cell cycle. Previous studies have identified v-cyclin D messenger RNA (mRNA) in biopsy specimens of Kaposi's sarcoma. In this study, we isolated a full-length v-cyclin D complementary DNA and characterized the pattern of v-cyclin D mRNA expression in Kaposi's sarcoma. METHODS: Standard methods were used to construct and to screen HHV-8 genomic and complementary DNA libraries. Reverse transcription-polymerase chain reaction (RT-PCR) methods and in situ hybridization with RNA probes were used to examine v-cyclin D mRNA expression. RESULTS: RT-PCR demonstrated the presence of v-cyclin D mRNA in biopsy specimens of AIDS-related Kaposi's sarcoma, in early-passage spindle cells from classical (i.e., not AIDS-related) Kaposi's sarcoma, and in spindle cells isolated from the peripheral blood of patients with AIDS-related Kaposi's sarcoma. In situ hybridization indicated that mRNAs for v-cyclin D and kaposin, an HHV-8 latency-associated gene, were present in approximately 1% of the spindle cells in early patch lesions and approximately 60% of the spindle cells in late nodular lesions of Kaposi's sarcoma. CONCLUSIONS: Spindle cells of Kaposi's sarcoma, which have been regarded as the tumor cells of this cancer, contain v-cyclin D mRNA. Expression of v-cyclin D protein may be involved in the pathogenesis of Kaposi's sarcoma by promoting cell proliferation.

[Risk factors of level â ¥ lymph node metastasis in cN0 papillary thyroid carcinoma].

Objective:To study the risk factors related to level Ⅵ lymph node metastasis in clinical N0 (cN0) papillary thyroid carcinoma (PTC). Method:A total of 107 cases with cN0 PTC treated in the same group were analyzed retrospectively. The frequency and risk factors for level Ⅵ lymph node metastasis in these patients were analyzed. Result:Level Ⅵ lymph node metastasis existed in 51.40% (55/107) cases. In univariate analysis, level Ⅵ lymph node metastasis was associated with age (χ²ï¼9.090,P<0.01), gender (χ²ï¼5.061,P<0.05), tumor maximum diameter (χ²ï¼8.772,P<0.01), tumor multifocality (χ²ï¼8.120,P<0.01), capsular invasion (χ²ï¼4.960,P<0.05), and surrounding tissue invasion (χ²ï¼3.858,P<0.05), but not with nodular goiter or Hashimoto's thyroiditis. Multivariate logistic analysis indicated that age,tumor maximum diameter, multifocal tumors and surrounding tissue invasion were independent risk factors for level Ⅵ lymph node metastasis. Conclusion:A high risk level Ⅵ lymph node metastasis exists in DTC with clinical N0. Prophylactic level Ⅵ neck dissection is strongly recommended in patients with PTC who are younger, tumor size more than 2 cm, multifocal tumor and surrounding tissue invasion.

Detection of human B-lymphotropic virus (human herpesvirus 6) sequences in B cell lymphoma tissues of three patients.

A survey for HBLV (human B-lymphotropic herpesvirus or human herpesvirus 6) sequences by Southern blot analyses was performed on DNA obtained from a variety of pathologically defined tissues, including a number of lymphomas, leukemias, and tissues from other hematologic disorders. Of the over 50 specimens studied, viral sequences were detected in three lymphomas of B cell derivation: an Epstein-Barr virus-positive African Burkitt's lymphoma, a follicular large cell lymphoma (nodular histocytic lymphoma), and two Epstein-Barr virus-negative tumors from a patient with Sjogren's syndrome. These results are the first indication of HBLV sequences associated with B cell tumors in a limited number of cases and raise the possibility that the virus might be involved in the genesis of some B cell tumors.

Novel organizational features, captured cellular genes, and strain variability within the genome of KSHV/HHV8.

Strong serologic and molecular probe correlations indicate that the newly discovered gamma herpesvirus KSHV or HHV8 is the likely etiologic agent of all forms of Kaposi's sarcoma as well as BCBL/PEL and MCD in patients with acquired immunodeficiency syndrome (AIDS). Two large segments of HHV8 DNA from an AIDS-associated BCBL tumor covering genomic positions 0-52 kilobase [kb] and 108-140 kb have been cloned, mapped, and partially sequenced. Our studies have focused on novel viral proteins encoded within a 13-kb divergent locus (DL-B) by nine captured homologues of cellular genes, including vIL-6, vDHFR, vTS, vBcl-2, three C-C beta chemokines (vMIP-1A, vMIP-1B, and vBCK), and two LAP/PHD subclass zinc finger proteins (IE1A and IE1B). The HHV-8 vIL-6, vDHFR, vTS, and vBcl-2 proteins have all been shown to be active in a variety of appropriate functional assays, and transcripts from vIL-6, vMIP-1B, vIE1-A, vIE1-B, and vDHFR genes are all expressed as abundant single messenger RNA species after butyrate or phorbol ester (TPA) induction of the lytic cycle in HHV8-positive BCBL cell lines. All of these genes lie within a divergent transcriptional domain that contains a single central enhancer and associated untranslated leader region plus seven distinct proximal promoters, some of which are negatively regulated through AP-1 and ZRE motifs by the EBV ZTA transactivator. This region also encompasses a predicted complex oriLyt domain of 1050 bp that is duplicated in inverted orientation adjacent to the T0.7 latency RNA in another large divergent locus (DL-E). We have previously described three distinct subtypes of the HHV8 genome that differ by 1.0%-1.5% at the nucleotide level within the ORF26 and ORF75 genes. Certain strains or clades appear to have preferential geographic distributions, but it is not known as yet whether there are any specific disease associations. Interestingly, the A, B, and C subtypes of HHV-8 also proved to differ dramatically in coding content at both the extreme left and right ends of the unique segment of the genome as well as in the positions of the junctions with the terminal repeats. On the left-hand side, the receptor-like ORF-K1 protein is highly variable with A-strain subtypes displaying 15% amino acid differences from C strains and up to 30% differences from B strains. On the right-hand side, two unrelated alternative types of the putative multiple membrane spanning ORF-K15 protein are found.

Structure of the long terminal repeat of simian lymphotropic virus type III (African green monkey) and its relatedness to that of HIV.

The simian T-lymphotropic virus type III (STLV-III[AGM]) is a retrovirus in wild African green monkeys which is serologically related to the human T-lymphotropic virus type III (HTLV-III/LAV-1/HIV) and other related human retroviruses. The long terminal repeats (LTR) contained in clones of viral DNA of (STLV-III[AGM]) were subcloned in M13 and their DNA sequence was determined and compared with that of HIV (HTLV-III[BH10]). The STLV-III(AGM) LTR is considerably larger than that of HTLV-III(BH10) (800 bp vs 634 bp) and contains a 498 bp U3 region, a 176 bp R region, and a 126 bp U5 region. These two LTR sequences share regions of significant homology. Regions of greatest homology include the 5' portion of U3, a core enhancer sequence in U3, sequences including and surrounding the TATAA promoter box in U3 and the AATAAA polyadenylation/termination signal in R, and the 3'-most region of U5. The relatively larger size of the STLV-III LTR is due to the presence in all three parts of the LTR of sequences which have no apparent homolog in the HIV LTR. Overall, the two LTRs are 47% homologous. Even greater homology (75%) is evident with a 300 bp segment including R and some of U3 from the LTR of another human retrovirus, HIV-2/LAV-2. The STLV-III LTR contains an imperfect 28 bp direct repeat in the R region which is not present in HIV. There are no obvious direct repeats in U3 homologous to the 10 bp repeat in the U3 of HTLV-III.(ABSTRACT TRUNCATED AT 250 WORDS)