Overexpression of connexin 40 (Cx40) inhibits the proliferation of H9c2 cardiomyocytes in rats by cell cycle arrest / 细胞与分子免疫学杂志

Objective To establish a stable strain of H9c2 cardiomyocytes overexpressing Cx40 and preliminarily investigate the effect of lentiviral vector-mediated Cx40 protein overexpression on the proliferation of H9c2 cells and its related mechanisms. Methods The Cx40 gene fragment was cloned from H9c2 cells by PCR and linked with lentivirus vector pLVX-IRES-Puro to obtain the recombinant plasmid pLVX-Flag-Cx40. Recombinant lentiviral particles carrying Flag-Cx40 were obtained by cotransfection with packaging plasmids into HEK293T cells. A stable expression strain (H9c2-Flag-Cx40 cell) was screened from infected H9c2 cells by purinomycin. The expression of Cx40 protein was detected by Western blot analysis, and the effect of Cx40 on H9c2 cells proliferation was determined by CCK-8 assay; cell cycle changes were measured by flow cytometry; the expression of the cell cycle protein cyclin D1 was detected by qRT-PCR and Western blot analysis. Co-immunoprecipitation (Co-IP) immunoprecipitation and Western blot analysis were used to identify the binding of Cx40 and Yes associated protein (YAP) in H9c2 cells; cytoplasmic and cytosolic proteins were isolated to detect the effect of Cx40 on the localization of YAP using Western blot analysis. Results Sequencing results showed that the recombinant pLVX-Flag-Cx40 expression vector was successfully established. A stable transfected cell line containing recombinant Flag-Cx40 lentivirus (H9c2-Flag-Cx40 cell) was successfully constructed from H9c2 cells. Compared with the control group, overexpression of Cx40 significantly reduced the proliferation of H9c2 cells, arrested the cell cycle at G0/G1 and reduced cyclin D1 expression. A significant increase in YAP expression was observed in the cytoplasm of the H9c2-Flag-Cx40 stable cell line, while the expression in the nucleus was significantly reduced. Cx40 bound to YAP in the cytoplasm and prevented it from entering the nucleus to play the role of transcriptional coactivation. Conclusion Overexpression of Cx40 induces cell-cycle arrest at G0/G1 phase and inhibits the proliferation in H9c2 cells.

Revision of childhood acute erythroid leukemia in the diagnosis by World Health Organization 2016 classification / 中华实用儿科临床杂志

Objective:To analyze the relationship between FAB morphological classification and World Health Organization (WHO) 2016 classification in children with acute erythroid leukemia(AEL), and to summarize the clinical features and prognosis.Methods:Clinical data of de nova childhood AEL patients from January 1, 2002 to December 31, 2019, in Pediatric Blood Disease Center, Institute of Hematology & Blood Disease Hospital were retrospectively analyzed.All of them were re-evaluated according to the WHO 2016 classification.Results:(1) A total of 20 patients were diagnosed as AEL by FAB classification.According to the criteria of WHO 2016, they were re-diagnosed as myelodysplastic syndromes (MDS)- refractory anemia with excess of blasts (11 cases), acute myeloid leukemia with MDS-related changes (3 cases), acute monocytic leukemia (1 case), and pure red leukemia (PEL, 5 cases). (2) Pathological hematopoiesis was frequently detected in bone marrow smears.Auer bodies were seen occasionally in some blasts.The most common antigen expressing were CD 117, CD 13, CD 33, CD 34, CD7, and CD 38.Karyotype analysis was performed in 18 cases successfully, involving 6 cases with abnormal karyotypes, including + 8, -7, 22p+ , t (3; 5: ? ), + 3q-, 15q-, and del (9)(q13). (3) Thirteen cases were treated by chemotherapy, and the one-course complete remission rate was 38.5%.By July 1, 2020, only 2 cases were alive without disease.The overall survival was 49 months and 11 months, respectively. Conclusions:Childhood AEL is susceptible to pathological hematopoiesis, poor response to early chemotherapy and poor prognosis.After re-evaluation according to WHO 2016 classification, most of them were diagnosed as MDS-related.Therefore, adjusting the suitable induction regimen with allogeneic hematopoietic stem cell transplantation may improve the prognosis.

Development of a biomechanical model for dynamic occlusal stress analysis / 国际口腔科学杂志·英文版

The use of traditional finite element method (FEM) in occlusal stress analysis is limited due to the complexity of musculature simulation. The present purpose was to develop a displacement boundary condition (DBC)-FEM, which evaded the muscle factor, to predict the dynamic occlusal stress. The geometry of the DBC-FEM was developed based on the scanned plastic casts obtained from a volunteer. The electrognathographic and video recorded jaw positional messages were adopted to analyze the dynamic occlusal stress. The volunteer exhibited asymmetrical lateral movements, so that the occlusal stress was further analyzed by using the parameters obtained from the right-side eccentric movement, which was 6.9 mm long, in the stress task of the left-side eccentric movement, which was 4.1 mm long. Further, virtual occlusion modification was performed by using the carving tool software aiming to improve the occlusal morphology at the loading sites. T-Scan Occlusal System was used as a control of the in vivo detection for the location and strength of the occlusal contacts. Data obtained from the calculation using the present developed DBC-FEM indicated that the stress distribution on the dental surface changed dynamically with the occlusal contacts. Consistent with the T-Scan recordings, the right-side molars always showed contacts and higher levels of stress. Replacing the left-side eccentric movement trace by the right-side one enhanced the simulated stress on the right-side molars while modification of the right-side molars reduced the simulated stress. The present DBC-FEM offers a creative approach for pragmatic occlusion stress prediction.

Audiological Outcomes of Sophonotron Bone Conduction Hearing Aid from a Multicenter Investigation / 听力学及言语疾病杂志

Objective The aim of the study was to evaluate the hearing performance of a new transcutaneous bone conduction hearing aid.Methods One hundred and nine patients with conductive or mix hearing loss and eleven patients with single sided sensorineural hearing loss from four tertiary referral centers were tested under unaided and aided conditions with the sound processors on testbands.Free field test was performed to obtain the hearing thresholds and speech recognition thresholds in quiet.Adverse events were also documented by a self-reported questionnaire.Results ①Conductive or mixed hearing loss patients:for aged≥6 yrs subgroup,the bone conduction PTA average of aided sides was 18.55±8.99 dB HL.For aged<6 yrs subgroup, the bone conduction ABR hearing threshold average was 18.33±8.36 dB HL.②Statistically significant improvements in free field hearing thresholds were seen when compared aided hearing with unaided outcomes.The average PTA (pure tone average in free field) was measured at 32.21±10.00, 37.33±14.15, and 34.38±10.76 dB HL respectively in conductive or mixed hearing loss aged≥6 yrs subgroup;aged<6 yrs subgroup and SSD subgroup.③In aided situations;speech recognition thresholds in quiet were significantly better compared to unaided situation in patients aged≥6yrs.④None of patients presented adverse events related to the devices.Conclusion The Sophono bone conduction hearing aid has satisfactory hearing performance and can be considered an alternative device for patients with conductive or mix hearing loss or with SSD.

The preparation of the mouse monoclonal antibodies specific for the DNAH2 protein / 天津医药

Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein,axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1-300 aa), in E. coli was constructed. IPTG was used to induce the expression of His-immunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal anti-DNAH2 antibody is prepared successfully.

Research progress on the anti-tumor differentiation effects of zoledronic acid in breast cancer / 天津医药

Breast cancer is the highest incidence and mortality of malignant tumor in women. Recurrence and distant metastasis are the main cause of death. Zoledronic acid (ZOL) has the potential to inhibit bone resorption characteristics mediated by osteoclast, which not only could suppress tumor cell proliferation and start the apoptosis of tumor cells, but also might interfere the adhesion of cancer cells to bone matrices, thereby could inhibit the migration and invasion of tumor cells. In some preclinical studies zoledronic acid has been demonstrated to have the direct anti-tumor effects on breast cancer. There are different therapeutic effects in treatment with zoledronic acid between the premenopausal and post-menopausal patients with breast cancer. This review summarized basic and clinical research progress of the anti-tumor differentiation effect of zoledronic acid in breast cancer.

The influence factors of the efficiency of semiconductor laser therapy for the treatment of temporoman-dibular disorders pain / 实用口腔医学杂志

Objective:To study the influence factors of laser therapy in the treatment of temporomandibular disorders(TMD)pain. Methods:1 20 cases of TMD were treated by MDC-500 semiconductor laser therapy.VAS was valued after each treatment.The cor-relation of the VAS varians following treatment with gender and age was analysed by two way AHOVA of the repeated measurement data.Results:1 1 4 cases completed the treatment and follow-up.After each treatment the VAS decreased(P <0.001 ),in females decreased more than in males(P <0.05).There was no significant difference between age groups(P =0.208).Conclusion:Laser therapy can ease TMD pain,and is more effective for female patients.

Dipsacus asperoides combined with composite calcium phosphate bone implant for bone defect repair / 中国组织工程研究

BACKGROUND:Physical structure and inorganic composition of composite calcium phosphate bone implants are similar to human bone, and the implants have good bioabsorbable ability and biocompatibility. Studies have confirmed that Dipsacus powder can significantly improve the speed of bone defect repair. OBJECTIVE:To study the effects of Dipsacus asperoides/composite calsium phosphate bone implant material in the repair of bone defects. METHODS:Bone defects of 1.0 cm×0.5 cm×0.3 cm were made on the bilateral mandibular bodies. The right side was implanted with Dipsacus asperoides/composite calsium phosphate bone implant as experimental group and the left side implanted with calcium phosphate bone material served as control group. Samples were col ected at 4, 8, 12 weeks after implantation for gross observation, cone beam CT, scanning electron microscope observation and histological observation. RESULTS AND CONCLUSION:(1) Gross observation:the osteogenic speed, degradation rate and hardness in the experimental group were higher than those in the control group. (2) Cone beam CT detection:the implant in the experimental group was more tightly integrated with the surrounding tissues than that in the control group, and the degradation speed in the experimental group was higher than that in the control group. (3) Scanning electron microscope observation:there were many fibrous connective tissues between the implants and the surrounding tissues in the two groups, which was more obvious in the experimental group than the control group. With time gone, the implants were combined with the surrounding tissues more tightly. (4) Histological observation:the osteogenic speed and activity in the experimental group were superior to those in the control group. These findings indicate that Dipsacus asperoides combined with composite calcium phosphate bone implant can accelerate bone repair.

Clinical examination of olfactory and gustatory function / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore changes of olfactory and gustatory functions in patients with olfactory dysfunction.@*METHOD@#The 284 study subjects included 92 healthy volunteers, 92 with hyposmia and 100 with functional anosmia. Their olfactory and gustatory functions were examined using T&T olfactometer, olfactory event-related potentials (OERPs) and triple drop method, respectively.@*RESULT@#The T&T results showed that the difference between patients with hyposmia and functional anosmia and healthy subjects had statistical significance. The OERPs results showed that patients with olfactory dysfunction had N1 and P2 waves of prolonged latency and reduced amplitude when compared to healthy subjects with the difference of statistical significance. When compared to healthy subjects, patients with olfactory dysfunction had clear hypogeusia and the difference had statistical significance. There was no significant difference between female and male groups.@*CONCLUSION@#It is suggested that the apparently concomitant hypogeusia is in patients with olfactory dysfunction. Decreased of olfactory and gustatory function exhibited little or no relationship with gender. Combined examinations of OERPs, T&T and triple drop method, could make it possible to reflect the overall chemical sensory functions in subjects systematically and provide scientific evidence for clinical diagnosis and treatment early.

Expression of herpesvirus-associated ubiquitin specific protease(HAUSP) in breast carcinoma and its clinical significance / 中国癌症杂志

Background and purpose: Herpesvirus-associated ubiquitin-specific protease (HAUSP) is a new deubiquitinating enzyme that was recently discovered. It has been demonstrated that HAUSP could deubiquitinate p53 both in vitro and in vivo. These results suggested that HAUSP might act as a tumor suppressor through the stabilization of p53 protein. The aim of this study was to investigate the expression of HAUSP in breast carcinoma, and its association with p53 protein as well as their relationship to prognosis. Methods: The expression of HAUSP mRNA was detected by real-time PCR. HAUSP protein and p53 protein were detected by immunohistochemistry with EnVision system in breast carcinoma tissues and noncancerous tissues. The relationship between their expressions and clinical pathological parameters were analyzed. Results: The expression of HAUSP mRNA was significantly lower in breast cancer tissue than noncancerous tissue (1.85±0.04 vs. 2.74±0.03, P<0.01). The positive rates of HAUSP protein were significantly lower in breast cancer tissue in noncancerous tissue (59.4% vs. 75.0%, P<0.01), and expression of HAUSP protein had no significant correlation with the clinical pathological parameters. There was no significant correlation between HAUSP and p53 protein (P>0.05). The expression of HAUSP mRNA was positively associated with that of HAUSP protein (P<0.01). There was no significant correlation between HAUSP and p53 protein. DFS of patients with both HAUSP positive and p53 negative was significant higher than the controls (P<0.01). Conclusion: Down-regulation of HAUSP protein and HAUSP mRNA in breast carcinoma indicated that HAUSP gene might correlated to tumor carcinogenesis. In addition, the simultaneous evaluation of both HAUSP expression and p53 expression status may be helpful to evaluate the prognosis of breast cancer patients.

The expression and significance of NF-kappaB in the rat model of otitis media with effusion experimentally induced by ovalbumin / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the T helper cells (Th) predominant differentiation and the modulation of nuclear transcription factors kappa B (NF-kappaB) in middle ear of rat model of otitis media with effusion (OME).@*METHOD@#Sixteen SD rats were randomly divided into OME (Exp group) and control group (Con group). The expression of NF-kappaB were observed by immunohistochemistry. The level of IFN-gamma and IL-4 in tympanic lavage fluid (TLF) were determined by ELISA.@*RESULT@#As compared to the Con group , the level of IL-4 and the ratio of Th2/Th1 (IL-4/IFN-gamma) in TLF of Exp group significantly increased (P<0.05), when no significant difference in IFN-gamma levels in TLF was found. The ratio of NF-kappaB p65 positive cells to white cells in temporal bone marrow smears and middle ear mucosa of Exp group was significantly higher than that of Con group (P<0.05). The expression of NF-kappaB p65 in temporal bone marrow smears and middle ear mucosa was signficantly positively correlated with the concentration of IL-4 in TLF of Exp group (P<0.05).@*CONCLUSION@#The middle ear is capable of mounting an allergic response and subsequent formation of effusion. There is Thl/Th2 immune response imbalance, which polarizes toward Th2 response in the middle ear microenvironment of allergic OME rat model. Moreover , NF-kappaB may participate in regulating Th2 predominant reaction.