Characterization of cellular senescence in doxorubicin-induced aging mice.

With a rise in the need to develop anti-aging drugs, a growing number of in vivo studies evaluating the efficacy of potential drug candidates have used doxorubicin-induced aging mice. However, changes in the biomarkers of senescent cells have not been reported in detail in these animals. To lay a foundation for the use of doxorubicin-induced aging mice, we examined the biomarkers of hepatic and renal senescent cells in these mice. We found that the 5 mg/kg doxorubicin dose is optimal to induce cellular senescence in mice. Subsequently, using this dose, we found that doxorubicin-induced an increase in senescence-associated ß-galactosidase (SA-ß-gal) positive cells in the kidney and lipofuscin accumulation in the liver. Some markers of senescent cells (p21WAF1/CIP1, p16INK4A, and γH2AX) were also significantly upregulated by doxorubicin and then counteracted by metformin treatment. These preliminary findings support the application of doxorubicin-induced aging mice as an animal model to evaluate the efficacy of anti-aging drug candidates.

Antiaging Effects of Vicatia thibetica de Boiss Root Extract on Caenorhabditis elegans and Doxorubicin-Induced Premature Aging in Adult Mice.

The roots of Vicatia thibetica de Boiss are a kind of Chinese herb with homology of medicine and food. This is the first report showing the property of the extract of Vicatia thibetica de Boiss roots (HLB01) to extend the lifespan as well as promote the healthy parameters in Caenorhabditis elegans (C. elegans). For doxorubicin- (Doxo-) induced premature aging in adult mice, HLB01 counteracted the senescence-associated biomarkers, including P21 and γH2AX. Interestingly, HLB01 promoted the expression of collagen in C. elegans and mammalian cell systemically, which might be one of the essential factors to exert the antiaging effects. In addition, HLB01 was also found as a scavenger of free radicals, thereby performing the antioxidant ability. Lifespan extension by HLB01 was also dependent on DAF-16 and HSF-1 via oxidative stress resistance and heat stress resistance. Taken together, overall data suggested that HLB01 could extend the lifespan and healthspan of C. elegans and resist Doxo-induced senescence in mice via promoting the expression of collagen, antioxidant potential, and stress resistance.

Cardiac metallothionein overexpression rescues diabetic cardiomyopathy in Akt2-knockout mice.

To efficiently prevent diabetic cardiomyopathy (DCM), we have explored and confirmed that metallothionein (MT) prevents DCM by attenuating oxidative stress, and increasing expression of proteins associated with glucose metabolism. To determine whether Akt2 expression is critical to MT prevention of DCM, mice with either global Akt2 gene deletion (Akt2-KO), or cardiomyocyte-specific overexpressing MT gene (MT-TG) or both combined (MT-TG/Akt2-KO) were used. Akt2-KO mice exhibited symptoms of DCM (cardiac remodelling and dysfunction), and reduced expression of glycogen and glucose metabolism-related proteins, despite an increase in total Akt (t-Akt) phosphorylation. Cardiac MT overexpression in MT-TG/Akt2-KO mice prevented DCM and restored glucose metabolism-related proteins expression and baseline t-Akt phosphorylation. Furthermore, phosphorylation of ERK1/2 increased in the heart of MT-TG/Akt2-KO mice, compared with Akt2-KO mice. As ERK1/2 has been implicated in the regulation of glucose transport and metabolism this increase could potentially underlie MT protective effect in MT-TG/Akt2-KO mice. Therefore, these results show that although our previous work has shown that MT preserving Akt2 activity is sufficient to prevent DCM, in the absence of Akt2 MT may stimulate alternative or downstream pathways protecting from DCM in a type 2 model of diabetes, and that this protection may be associated with the ERK activation pathway.

Effects of Breast Cancer Genes 1 and 2 on Cardiovascular Diseases.

Carriers of mutations of breast cancer gene 1 and/or 2 (BRCA1/2) have a higher risk of developing breast and ovarian cancers at a relatively young age. Recently, a causative role for BRCA1/2 in cardiovascular diseases has been emerging. In this review, we summarize currently available evidence obtained from studies on animal models and human BRCA1/2 mutation carriers that shows a correlation of BRCA1/2 deficiency with various cardiovascular diseases, including ischemic heart disease, atherosclerosis, and chemotherapy-linked cardiac muscle disorders. We also discuss one of the major mechanisms by which BRCA1/2 protects the heart against oxidative stress, ie mediating the activity of Nrf2 and its downstream targets that govern antioxidant signaling. More research is needed to elucidate whether the carriers of the BRCA1/2 mutations with ovarian and breast cancers have increased susceptibility to chemotherapy-induced cardiac functional impairment.

Cardiac injury prediction and lymphocyte immunity and inflammation analysis in hospitalized patients with coronavirus disease 2019 (COVID-19).

BACKGROUND: Coronavirus disease 2019 (COVID-19) is an ongoing global pandemic. The ability to predict cardiac injury and analyze lymphocyte immunity and inflammation of cardiac damage in patients with COVID-19 is limited. We aimed to determine the risk factors and predictive markers of cardiac injury in these patients. METHODS: Data from 124 consecutive hospitalized patients with confirmed COVID-19 were collected. We compared the proportion of cardiovascular disease history in moderate, severe, and critical cases. We obtained high-sensitivity cardiac troponin I (hs-cTn I) results from 68 patients. Patients were divided into two groups based on positive hs-cTn I result: those with cardiac injury (n = 19) and those without cardiac injury (n = 49). RESULTS: Compared with the group with moderate disease, hypertension, coronary heart disease, and smoking were more common in severe and critical cases. Diabetes mellitus was most common in the critical group. Age older than 65 years, presence of chronic kidney disease, and lower blood lymphocyte percentage were independent risk factors of cardiac injury. The total T- and B-lymphocyte counts and CD4+ and CD8+ T-cell counts were significantly lower in those with cardiac injury. A minimal lymphocyte percentage < 7.8% may predict cardiac injury. The interleukin (IL) 6 level in plasma was elevated in the group with cardiac injury. CONCLUSIONS: The lymphocyte percentage in blood may become a predictive marker of cardiac injury in COVID-19 patients. The total T and B cells and CD4+ and CD8+ cell counts decreased and the IL-6 level increased in COVID-19 patients with cardiac injury.

[Nasal bleeding as the first symptom of tsutsugamushi disease: a case report].

A case of 76-years-old male patient with nasal bleeding as the first symptom in our hospital, who was finally diagnosed as tsutsugamushi disease. This old man was bited by insect in farmland 2 days before the symptom occurred. PE: Left thigh and right buttock have eschar, with splenomegaly. Routine blood test: WBC (decrease) 3.9 x 10(9)/L, RBC (decrease) 3.86 x 10(9)/L, PLT (decrease) 41 x 10(9)/L, HGB (decrease) 117 g/L; Chest CT: lung interstitial pneumonia, a small amount of bilateral pleural effusion. Oxk-ag 1:320. The patient was discharged after treatment with chloramphenicol for 8 days.

[Correlation of VEGF expression with microvessel and microlymphatic density in sinonasal squamous cell carcinoma].

OBJECTIVE: To study the expression of VEGF in sinonasal squamous cell carcinoma and its correlations with microvessel density (MVD), microlymphatic vessel density (MLVD). METHOD: The expression of VEGF, MVD and MLVD in 41 cases of sinonasal squamous cell carcinoma were detected by immunohistochemical technique. RESULT: In the sinonasal squamous cell carcinoma, the positive rate of VEGF was 82.9% (34/41). The over expression of VEGF was related with tumor invasion, histological grading and lymphatic metastasis (P < 0.05). The MVD of cases with positive VEGF expression was significantly higher than those without VEGF expression (P < 0.05), but was not statistical difference in MLVD (P > 0.05). CONCLUSION: VEGF may participate in the metastasis of sinonasal squamous cell carcinoma through promoting vascularization in the tumors, but not promoting MLVD.

[The significance of the expression of Col IV and LN in nasal and paranasal sinus malignant tumor].

OBJECTIVE: To study the pathological relationship between the expression of Col IV and LN in nasal and paranasal sinus malignant tumor (NPMT). METHOD: The immunohistochemical technique was used to detected the expression of Col IV and LN in NPMT, para-cancer tissues and non-cancer tissues. RESULT: There was a significance on the expression of the Col IV and LN in NPMT, para-cancer tissues and non-cancer tissues (P<0.01), and no significance in endepidermis and soft connective tissue of the NPMT (P>0.05). CONCLUSION: The Col IV and LN perhaps participate in tumorigenesis of NPMT, and may play the homoioplastic role in different pathological types of the NPMT.

[Study about eye complication of nasopharyngeal carcinoma].

OBJECTIVE: To explore the eye complication of nasopharyngeal carcinoma (NPC), to analysis the clinical manifestation, CT characteristics and pathological diagnosis of eye complications of NPC and to provide the base for early diagnosis of NPC. METHOD: To retrospectively study of 82 cases eye complications in 562 cases NPC, to study their clinical manifestation, CT characteristics and pathological diagnosis. RESULT: The clinical studies showed that eye complication cases were occurred in 82 cases of 562 NPC cases (14.6%). Thirty-six cases in left and 37 cases in right eye, 9 cases in bilateral eyes. Sixty-five cases came from Guangdong, while the others 17 cases come from 5 provinces. There were 9 kinds of eye manifestation. CT appearances: 40 cases suffered from skull base distracted, 6 cases with orbit involved, 2 cases ( in left eyes) with orbit metastasis, 12 cases with nose-sinus involved, 68 case with parapharyngeal space involved, 49 cases with soft issue in wall of styloid process involved (there were many kind of shows in the same case, so the data were repeated in these cases). CONCLUSION: There were multiplicity and complexity in eye complication of NPC. Ophthalmologists should think highly of these cases.

[Cluster analysis in micrangium detection in malignant nasal and paranasal sinus tumor].

OBJECTIVE: To study the application of cluster analysis in micrangium detection in malignant nasal and paranasal sinus tumor. METHODS: Microvessel density (MVD) counting and cluster analysis were used to detect the micrangium in patients with malignant nasal and paranasal sinus tumor to assess the association between the malignancy and MVD. RESULTS: According to cluster analysis, the MVD counting could be clustered into two groups, and the MVD showed significant differences between the tumor tissues, adjacent normal tissue and the control group (P<0.01), a result consistent with that by analysis of variance of the MVD. CONCLUSION: Cluster analysis can be used in clustering of MVD counting in malignant nasal and paranasal sinus tumor to simplify MVD counting, and offers an important analytic method for micrangium analysis in tumors.

[Inhibition of growth and proliferation of Hep-2 cells by targeting hTERT gene using small interference RNA technology].

OBJECTIVE: To investigate the inhibition of hTERT gene to Hep-2 cells of laryngeal neoplasms. METHOD: siRNA-hTERT mRNA was designed. Hep-2 cells, cultured in vitro, were transfected with lipofectamine 2000 and the inhibitory effect was detected by reverse transcriptase (RT-PCR), MTT assay, and morphology. RESULT: 1. siRNA were transfected into Hep-2 cells successfully; 2. RT-PCR showed the exogenous hTERT was knocked down 70% by siRNA; 3. The morphology and the MTT result showed the hTERT siRNA to be able effectively to suppress the Hep-2 cell multiplication, the suppression rate achieves 65%. CONCLUSION: siRNA target to hTERT can remarkably suppress the proliferation of Hep-2 cell.

[Inhibition of growth and proliferation of Hep-2 cells by targeting c-myc gene using small interference RNA technology].

OBJECTIVE: To investigate the effect of small interfering RNA (siRNA) targeting c-myc gene in Hep-2 cells. METHOD: siRNA targeting c-myc mRNA was designed and synthesized. In vitro cultured Hep-2 cells were transfected with lipofectamine 2000 and the inhibitory effect was detected by MTT, morphology, real time PCR assay. RESULT: 1) The MTT result showed the c-myc siRNA to be able effectively to suppress the Hep-2 cell multiplication; 2) The real time PCR result showed c-myc at mRNA level inhibition ratio at 94% in group S3; 3) The morphology result showed the c-myc siRNA to be able effectively to suppress the Hep-2 cell multiplication, the cell heteromorphism was diminished. CONCLUSION: siRNA targeting c-myc can remarkably suppress the Hep-2 cell growth and multiplication.

[Clinical significance of the expression of p53, p63 and p73 in nasal and paranasal sinus carcinoma].

OBJECTIVE: To study the clinical significance of the expression of p53, p63 and p73 protein and the correlation of p53 and p63 in nasal and paranasal sinus carcinoma (NPSC). METHOD: The immunohistochemical technique was used to detect the expression of p53, p63 and p73 in 67 cases of NPSC, para-cancer tissues and non-cancer tissues. RESULT: The positive rate of p53 and p63 protein in NPSC was significantly higher than that in park cancer tissues and non-cancer tissues (P<0.01), and there were a positive correlation between the expression of p53 and p63 protein in NPSC (P<0.01). But the expression of p73 had no significant difference among NPSC, para-cancer tissues and non-cancer tissues (P>0.05). CONCLUSION: There are positive correlation between p53 and p63 protein in NPSC, and they may play an important role in the tumorgenesis of NPSC. The p73 may not be associated with tumorgenesis of NPSC.

[Recombinant adeno-associated virus conducted NgR(DN) enhances axonal regeneration of optic nerve after trauma: experiment with rats].

OBJECTIVE: To investigate the effect of recombinant adeno-associated virus conducted NgRDN on the axonal regeneration of optic nerve after trauma. METHODS: Two kinds of adeno-associated virus (AAV), AAV-NgRDN-EGFP containing dominant negative form of Nogo receptor and enhanced green fluorescent protein (EGFP) and rAAV-NgR-EGFP containing Nogo-66 receptor (NgR) and EGFP, were constructed. 45 adult Wistar male rats were randomly divided into three equal groups, all with both eyes as experimental eyes: Groups A, B, and C to undergo injection of rAAV-EGFP, rAAV-NgR-EGFP, and rAAV-NgRDN-EGFP respectively into the vitreous; and each group was subdivided into 3 equal subgroups: subgroups 1 underwent injection of rAAV only, subgroups 2 underwent injection of rAAV and lens trauma, and subgroups 3 underwent injection of rAAV and zymosan. The rats in the Subgroups A2, B2, and C2 underwent. Crush of the optic nerve 2 mm behind the eyeball with optic nerve forceps 3 weeks after the injection. Four days after the crush the right eyes were taken out and the retinal explants were cultured in 2 kinds of culture fluid: with or without myelin. The growth of axons at the edge of retinal explants was observed by immunofluorescent staining with betaIII tubulin. Two weeks after the crush the other eyes were taken out to isolate the optic nerves. Immunofluorescence assay was used to detect the expression of growth associated protein-43 (GAP-43) of optic nerve. The axonal regeneration of optic nerve was observed. RESULTS: betaIII tubulin staining showed that on the condition of culture fluid without myelin both rAAV-NgR-EGFP and rAAV-NgRDN-EGFP showed no effects on the axonal regeneration of retinal ganglion cells (RGCs). However, on the condition of culture fluid with myelin the count of axonal regeneration and the length of regenerated axons of Group B were (13+/-4) and (36 microm+/-4 microm), both significantly lower than those of Group A [(21+/-4) and (83 microm+/-11 microm) respectively, both P<0.01]. There were not significant differences in count of axonal regeneration and length of regenerated axons between Subgroups C1 and A1. The count of axonal regeneration and length of regenerated axons of Subgroups C2 were (317+/-45) and (508 microm+/-44 microm), both significantly higher than those of Subgroup C3 [(238+/-30) and (365 microm+/-48 microm) respectively, both P<0.01], and the values of both Subgroups C2 and C3 were significantly higher than those of Subgroups A2 and A3. The GAP43-positive area in the optic nerve of Group C was significantly larger than that of Group A (P<0.01), and that of Group B was significantly smaller than that of Group A (P<0.01). The GAP43-positive area in the optic nerve of Subgroup A2 was (18.71+/-1.72)x100 microm2, significantly larger than that of Subgroup A3 [(12.75+/-1.02)x100 microm2, P<0.01], and that of Subgroup A3 was significantly larger than that of Subgroup A1 (P<0.01). There were not significant differences in the GAP43-positive area among the subgroups in Group B. CONCLUSION: Transfection of rAAV-NgRDN-EGFP into RGC in an activated status enhances axonal regeneration of optic nerve. NgRDN AAV can inhibit effectively the role of NgR.

[The clinical significance of the expression of p63 gene in laryngeal squamous cell carcinoma].

OBJECTIVE: To study the expression and clinical significance of p63 gene in laryngeal squamous cell carcinoma(LSCC). METHOD: The immunohistochemical technique was used to detected the expression of p63 in 63 cases of LSCC, 45 cases of para-cancer tissues, 24 cases of lymph node metastasis, 40 cases of non lymph node metastasis and 25 normal tissues. RESULT: The positive overexpression rate of the p63 gene in LSCC (95.24%) and in matched metastatic lymph node (83.3%) was significantly higher than para cancer tissues, non metastasis lymph node and normal tissues (P<0.01). p63 gene expression was not correlation with the clinical stages,clinical typing and pathological classification of the LSCC(P >0.05). CONCLUSION: p63 gene had important clinic significance to the diagnosis and differential diagnosis of the LSCC. It perhaps participate in regulation of tumorigenesis in LSCC, and it may play negative feedback action in the development of LSCC.

[Expression and clinical significance of matrix metalloproteinase-2 in laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the relationship between the expression of matrix metalloproteinase-2 (MMP-2) and clinical significance of laryngeal squamous cell carcinomas (LSCC). METHOD: Expression of the MMP-2 was detected by immunohistochemical method in 63 cases of LSCC, 20 cases of adjacent safety margin (ASM) and 18 cases of vocal cord polyp (VCP). RESULT: The positive percentage of active MMP-2 expression were 66.7%, 33. 3% and 15. 0%, in LSCC,VCP and ASM respectively. Significant differences ( P <0. 05) were observed among the three group; there was a significant differences in the expression of MMP-2 among the different pathologic degree I , II , III; no significant difference in the expression of MMP-2 between lymphoid metastasis and no lymphoid metastasis ( P >0. 05); Age and clinical stage did not change the expressions of the MMP-2 expression ( P <0. 05). CONCLUSION: MMP-2 probably plays an important role in the carcinogenesis and progression of LSCC. But it may be an unnecessary factor in the metastases of LSCC.

[Risk factors and preventive steps for nasosinusitis after radiotherapy of nasopharyngeal carcinoma].

OBJECTIVE: To study the risk factors and preventive steps of nasosinusitis after radiotherapy of nasopharyngeal carcinoma (NPC). METHOD: Two hundreds and thirtyone NPC patients were retrospectively analysed. RESULT: Among 231 NPC patients, 98 patients (42.4%) had nasosinusitis before radiotherapy, 165 patients (66.7%) suffered from nasosinusitis after radiotherapy. The rate of nasosinusitis in NPC patients with high T stage and nasalcavity anatomical aberrance was higher than in others (P = 0.014, CR = 1. 973; P = 0. 034, OR = 2.156), and the rate of nasosinusitis in NPC patients with nasopharynx therapy was low (P = 0.008, OR = 0.675), most patients with nasosinusitis before radiotherapy had not any improve after radiotherapy,and effected the effect of radiotherapy. CONCLUSION: To reduce nasosinusitis after radiotherapy and improve patients quality of life, we suggest that nasopharyngeal cavity therapy should be carried out during and after irradiation for NPC patients, especially for those patients with high T stage and nasalcavity anatomical aberrance, and for those who had nasosinusitis before radiotherapy.

[Expression and significance of PTEN and Survivin between the primary foci and the lymph node metastases foci of laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the molecular difference and significance between the primary foci and metastases foci of laryngeal squamous cell carcinoma (LSCC). METHOD: Tissues of 18 cases of positive lymph node metastases (N+) LSCC were detected by using SP method. The LSCC primary foci and metastases foci and negative metastases lymph nodes were included, 22 cases of vocal cord polyp (VCP) were regarded as control. expression of PTEN and Survivin were evaluated by SP immunohistochemistry. RESULT: Expression of neither PTEN nor Survivin were positive in negative nodes. The positive rates of PTEN in LSCC, N+, VCP were 94.4% (17/18), 94.4% (17/18) and 95.5% (21/22) respectively, there were no significant difference among them (P>0.05). But the expression degree were more powerful in N+ than in the LSCC (P<0.05). There was no expression of Survivin in VCP. The positive rates of Survivin in LSCC and N+ were 55.5% (10/18), 50.0% (9/18), the difference were significant (P<0.01); but between LSCC and N+, neither the positive rate nor the expression degree had significant difference (P>0.05). CONCLUSIONS: There were differences between the LSCC primary foci and the lymph node metastases foci in molecular level probably.

[Expression and clinical significance of p185 and CD44v6 in laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the expression and the clinical significance of proto-oncogene cerbB-2 proteins p185 and adhesion molecules CD44v6 in laryngeal squamous cell carcinomas (LSCC) and the relationship between them. METHOD: expression of p185 and CD44v6 was studied by immunohistochemical analysis from 63 cases of LSCC, 20 cases of adjacent safety margin (ASM) and 18 cases of vocal cord polyp(VCP). RESULT: There were no expression of p185in VCP, Significant differences (P < 0.05) were observed between the two group; there was significant differences among the different pathologic degree I, II, III; The expression of p185 was not related to age and clinical stage. Significant differences (P < 0.05) were found among LSCC, VCP and ASM; there was no differences between the different pathologic degree I, II, III. The expression of CD44v6 was not related to age and clinical stage. Significant correlation was observed between the expression of p185 and CD44v6 in LSCC. CONCLUSION: p185 probably plays an important role during the carcinogenesis and progression process of LSCC. While CD44v6 has the important function only in the carcinogenesis process of LSCC. The expression of p185 and CD44v6 were higher in ASM than in VCP. It indicates that p185 and CD44v6 were probably the unstable factor in ASM which related to the recurrent of laryngeal carcinoma, and they may cooperate with each other.